TRANSGENIC ANIMALS

OBJECTIVE: Animal biotechnology will !o"i#e com elling an# !actical bene$it% to man&in#' a% we ha"e %een $!om othe! $(n#amental a#"ance% in li$e %cience) Genetic enginee!ing i% the #elibe!ate mo#i$ication o$ the animal*% genome (%ing techni+(e% o$ mo#e!n biotechnology) Genetically enginee!e# ag!ic(lt(!al animal% a!e being #e"elo e# to t!an%$o!m an# im !o"e (blic health) Some ob,ecti"e% a!e gi"en below -) Genetically enginee!e# animal% will im !o"e h(man health th!o(gh #i%ea%e) .) Animal% that a!e genetically enginee!e# will ha"e im !o"e# $oo# !o#(ction t!ait% enabling them to hel meet the global #eman# $o! mo!e e$$icient' highe! +(ality an# lowe!/co%t %o(!ce% o$ $oo#) !o#(ction o$ no"el

!e lacement !otein%' #!(g%' "accine% an# ti%%(e% $o! the t!eatment an# !e"ention o$ h(man

0) Genetically enginee!e# animal% will cont!ib(te to im !o"ing the en"i!onment an# h(man health with the con%(m tion o$ $ewe! !e%o(!ce% an# the !o#(ction o$ le%% wa%te) 1) Genetic enginee!ing o$$e!% t!emen#o(% bene$it to the animal by enhancing health' well/being an# animal wel$a!e) 2) Genetically enginee!e# animal% ha"e !o#(ce# high/"al(e in#(%t!ial !o#(ct% %(ch a% % i#e! %il& (%e# $o! me#ical an# #e$en%e (! o%e%)

INTRO34CTION

INTRO34CTION: The!e a!e "a!io(% #e$inition% $o! the te!m t!an%genic animal) The 5e#e!ation o$ E(!o ean Labo!ato!y Animal A%%ociation% #e$ine% the te!m a% an animal in which the!e ha% been a #elibe!ate mo#i$ication o$ it% genome' the genetic ma&e( inhe!ite# cha!acte!i%tic%) 65ELASA -78.9 In the ea!ly -7:;%' (nea%e % !ea# th!o(gh the me#ia abo(t a new %cienti$ic techni+(e calle# !ecombinant #eo<y!ibon(cleic aci# 63NA9) The conce t wa% ea%y to (n#e!%tan#: yo( ta&e a gene o(t o$ one li"ing thing an# (t it in anothe!) =hen %cienti%t% !o o%e# to in%e!t h(man gene% into bacte!ia' whe!e they co(l# be mo!e ea%ily mani (late#' o %ocial an# %cienti$ic im lication%) Be$o!e thi% technology came along' $(n#amental a#"ance% on cell(la! #i%ea%e #i#n>t %eem o%%ible) Recombinant 3NA change# all that' an# in a %ho!t time' ga"e !i%e to new me#icine% an# in%ight% into many common #i%ea%e%) ?et in the -7:;%' %ome oll% %(gge%te# many onent% wo!!ie# abo(t (n$o!e%een o$ an o!gani%m !e% on%ible $o!

Ame!ican% we!e again%t the !e%ea!ch' ca ti"e to conce!n% abo(t it% e!cei"e# !i%&% an# willing to $o!go ob"io(% (blic health o o!t(nitie%) @!ominent c!itic% o$ the technology we!e con"ince# that A!ecombinantB bacte!ia we!e (n%a$e an# ca able o$ in$ecting eo le) =hen they !o o%e# a mo!ato!i(m on $(!the! !e%ea!ch' %ome B!iti%h !e%ea!che!% mi<e# the !ecombinant bacte!ia into thei! mil& an# #!an& it with no ill e$$ect%) The oint wa% ma#e) The mo!ato!ia ne"e! a%%e#) An# me#ical !actice ha% been t!an%$o!me# a% a !e%(lt) 6Scott Gottlieb .;;.9 A t!an%genic animal i% one who%e genome ha% been change# to ca!!y gene% $!om othe! % ecie%) The n(cle(% o$ all cell% in e"e!y li"ing o!gani%m contain% gene% ma#e ( o$ 3NA) The%e gene% %to!e in$o!mation that !eg(late% how o(! bo#ie% $o!m an# $(nction) Gene% can be alte!e# a!ti$icially' %o that %ome cha!acte!i%tic% o$ an animal a!e change#) 5o! e<am le' an emb!yo can ha"e an e<t!a' $(nctioning gene $!om anothe! %o(!ce a!ti$icially int!o#(ce# into it' o! a gene int!o#(ce# which can &noc& o(t the $(nctioning o$ anothe! a!tic(la! gene in the emb!yo) Animal% that ha"e thei! 3NA mani (late# in thi% way a!e &now% a% t!an%genic animal%) 6T!an%genic Animal% .;;09 The ma,o!ity o$ t!an%genic animal% !o#(ce# %o $a! a!e mice' the animal that ionee!e# the technology) The $i!%t %(cce%%$(l t!an%genic animal wa% a mo(%e) A $ew yea!% late!' it wa% $ollowe# by !abbit%' ig%' %hee ' an# cattle) 6Re<!oa# et al)-7879 6Ro%chla( -7879 6@(!%el et al)-7879 6hamme! et al)-7829 T!an%genic animal% a!e (%e$(l a% #i%ea%e mo#el% an# !o#(ce!% o$ %(b%tance% $o! h(man wel$a!e)

Some t!an%genic animal% a!e !o#(ce# $o! % eci$ic economic t!ait%) 5o! e<am le' t!an%genic cattle we!e c!eate# to !o#(ce mil& containing a!tic(la! h(man !otein%' which may hel in the t!eatment o$ h(man em hy%ema)

Othe! t!an%genic animal% a!e !o#(ce# a% #i%ea%e mo#el% 6animal% genetically mani (late# to e<hibit #i%ea%e %ym tom% %o that e$$ecti"e t!eatment can be %t(#ie#9) 5o! e<am le' Ca!"a!# %cienti%t% ma#e a ma,o! %cienti$ic b!ea&th!o(ghh when they !ecei"e# a 4)S) atent 6the com any 3(@ont hol#% e<cl(%i"e !ight% to it% (%e9 $o! a genetically enginee!e# mo(%e' calle# OncoMo(%e o! the Ca!"a!# mo(%e' ca!!ying a gene that !omote% the #e"elo ment o$ "a!io(% h(man cance!%) 6@hili Le#e!: Vignette% .;;09

Genetically enginee!e# animal% D which o$ten inco! o!ate gene% $!om othe! o!gani%m% in a !oce%% calle# t!an%gene%i% D a!e being #e"elo e# to t!an%$o!m an# im !o"e (blic health) The b!oa# o%%ibilitie% encom a%% the t!eatment o$ h(man #i%ea%e' the !o#(ction o$ %a$e! o! mo!e e$$ecti"e h(man !otein%' new #!(g% an# "accine%' the ea%ing o$ %ho!tage% o$ h(man ti%%(e an# o!gan% a"ailable $o! t!an% lant atient% th!o(gh new a"en(e% o$ %( ly' the enhancement o$ the en"i!onment an# %(%taining $oo# %ec(!ity an# +(ality th!o(gh the im !o"e# e$$iciency o$ $oo# !o#(ction' an# !o#(ction o$ mo!e n(t!itio(% $oo#%) 65(lton .;;;9 6Echela!# -77E9 6?o(ng et al -7789 6Reggio et al) .;;.9) The c!eation o$ the $i!%t genetically enginee!e# $a!m animal% wa% #oc(mente# in -782 an# the ca ability $o! bio ha!mace(tical t!an%genic !o#(ction by the%e animal% wa% #emon%t!ate# %ho!tly animal a lication% with bene$icial the!ea$te!) To#ay' the!e a!e mo!e than two #oFen #!(g% in #e"elo ment #e!i"e# th!o(gh metho#%' an# n(me!o(% ag!ic(lt(!al en"i!onmental an# h(%ban#!y att!ib(te% %(itable $o! comme!cialiFation) 6Camme! et al) -7829 @!io! to the #e"elo ment o$ molec(la! genetic%' the only way o$ %t(#ying the !eg(lation an# $(nction o$ mammalian gene% wa% th!o(gh the ob%e!"ation o$ inhe!ite# cha!acte!i%tic% o! % ontaneo(% m(tation%) Long be$o!e Men#el an# any molec(la! genetic% &nowle#ge' %electi"e b!ee#ing wa% a common !actice among $a!me!% $o! the enhancement o$ cho%en t!ait%' e)g)' inc!ea%e# mil& !o#(ction) 3(!ing the -7:;%' the $i!%t chime!ic mice we!e !o#(ce# 6B!in%te!' -7:19) The cell% o$ two #i$$e!ent emb!yo% o$ #i$$e!ent %t!ain% we!e combine# togethe! at an ea!ly %tage o$ #e"elo ment 6eight cell%9 to $o!m a %ingle emb!yo that %(b%e+(ently #e"elo e# into a chime!ic a#(lt' e<hibiting cha!acte!i%tic% o$ each %t!ain)

The m(t(al cont!ib(tion% o$ #e"elo mental biology an# genetic enginee!ing e!mitte# !a i# #e"elo ment o$ the techni+(e% $o! the c!eation o$ t!an%genic animal%) 3NA mic!oin,ection' the $i!%t techni+(e to !o"e %(cce%%$(l in mammal%' wa% $i!%t a lie# to mice 6Go!#on an# R(##le' -78-9 an# then to "a!io(% othe! % ecie% %(ch a% !at%' !abbit%' %hee ' ig%' bi!#%' an# $i%h) Two othe! main techni+(e% we!e then #e"elo e#: tho%e o$ !et!o"i!(%/me#iate# t!an%gene%i% 6Jaeni%ch' -7:E9 an# emb!yonic %tem 6ES9 cell/me#iate# gene t!an%$e! 6Go%%le! et al)' -78E9) Since -78-' when the te!m t!an%genic wa% $i!%t (%e# by J)=) Go!#on an# 5)C) R(##le 6-78-9' the!e ha% been !a i# #e"elo ment in the (%e o$ genetically enginee!e# animal% a% in"e%tigato!% ha"e $o(n# an inc!ea%ing n(mbe! o$ a lication% $o! the technology) TRANSGENIC ANIMAL DISEASE MODELS Mo!e than 0;;; h(man genetic #i%ea%e% a!e &nown' an# the!e i% m(ch inte!e%t in %t(#ying thei! $(n#amental ca(%e% %o that e$$ecti"e t!eatment% an# %omatic cell gene the!a ie% can be #e"i%e#) S eci$ic inb!e# mo(%e %t!ain%' which inhe!it % ontaneo(%ly #e!i"e# henoty e%' ha"e !o"i#e# (%e$(l mo#el% $o! in"e%tigating the animal%: athogene%i% o$ h(man #i%ea%e%) Ne"e!thele%%' %e"e!al !oblem% a!e a%%ociate# with %t(#ying nat(!ally occ(!!ing h(man genetic #i%ea%e% by (%ing

a9 Animal %t!ain% %howing e< en%i"e to maintainG

a!tic(la! #i%ea%e %ym tom% a!e o$ten #i$$ic(lt to obtain an#

b9 Thei! % eci$ic genetic #e$ect% can be a% #i$$ic(lt to i#enti$y an# cha!acte!iFe a% tho%e o$ thei! h(man co(nte! a!t%G an# c9 A$$ecte# animal% o$ten #i$$e! $!om (na$$ecte# cont!ol% in genetic $acto!% a##itional to the gene in +(e%tion)

ADVANTAGES O"e! the la%t #eca#e' many t!an%genic an# &noc&o(t m(tant mo(%e %t!ain% ha"e been c!eate# a% #i%ea%e mo#el%) Mo#el% e<i%t $o! ne(!o %ychiat!ic' ca!#io"a%c(la!' (lmona!y' oncological' in$lammato!y an# imm(nological #i%ea%e%' a% well a% $o! %t(#ying mechani%m% an# #i%o!#e!% o$ h(man metaboli%m' !e !o#(ction an# ea!ly #e"elo ment) The%e mo#el% a!e #oc(mente# in t!an%genic #ataba%e%' %(ch a% TBASE o! In#(ce# M(tant Re%o(!ce 6IMR9) So $a!' mainly %ingle/ gene con#ition% ha"e been %t(#ie#' b(t otential a lication% o$ t!an%genic mice incl(#e the analy%i% o$ olygenic #i%ea%e%) 6Be#ell et al)-77:9 6=oychic& et al)-7709

TRANSGENIC PROCESS
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TRANSGENIC @ROCESS:

Table I: Metho#% $o! !o#(cing t!an%genic animal% by int!o#(ction o$ $o!eign 3NA into the mammalian genome

Technique involved

Remark

@!on(clea! gene9

mic!o

in,ection technical %im licityG low %(cce%% !ateG a licable o$ e< !e%%e# to a wi#e !ange o$ % ecie%G mo%t wi#ely (%e#G (n !e#ictable e$$ect% #(e to !an#om t!an%gene integ!ation

6int!o#(ction !ange

Emb!yonic mani (lation

%tem

6ES9

cell %(b%tit(tion o$ a $(nctional gene with an inacti"e o$ geneG % ecie%' ge!m/line com etent ES cell% ha"e been incl(#ing !imate%' b(t toti otency

6int!o#(ction

e< !e%%e# gene' o! gene inacti"ation i%olate# in miceG ES/li&e cell% i#enti$ie# in othe! by homologo(% !ecombination9 !emain% to be e%tabli%he#

C!e/lo< techni+(e

!e$e!!e# metho# with mo!e cont!ol o"e! !e%(lting henoty eG time/con%(ming

Vi!al "ecto!%

com le<G la!gely !e%t!icte# to a"ian % ecie%

Cyto la%mic in,ection

le%% e$$icient than #i!ect !on(clea! mic!oin,ection

@!imo!#ial ge!m cell%

chimae!ic animal% !e%(lt

N(clea! t!an%$e!

la!ge otential $o! genetically mo#i$ying li"e%toc&

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S e!matogonial mani (lation

t!an% lantation into !eci ient te%te%

@!o#(ction o$ t!an%genic animal% able to t!an%mit genetic mo#i$ication% to thei! o$$% !ing !e+(i!e% ge!mline t!an%$o!mation) The!e a!e %e"e!al techni+(e% $o! the !o#(ction o$ t!an%genic animal% 6Table% I9G the th!ee mo%t commonly (%e# metho#%' !on(clea! mic!oin,ection' !et!o"i!al "ecto!an# the emb!yonic %tem 6ES9 cell mani (lation) T!an%genic animal can be !o#(ce# by th!ee metho#%: -) Mic!oin,ection .) Ret!o"i!al "ecto! 0) Emb!yonic %tem cell !" Microin#ec$ion% Thi% metho# wa% $i!%t #e"elo e# in -7EE) The mo(%e wa% the $i!%t animal to (n#e!go %(cce%%$(l gene t!an%$e! (%ing 3NA mic!oin,ection) Mic!oin,ection a%%embly con%i%t% o$ two mic!omani (lato!%' one $o! a gla%% mic!o i ette to hol# the o"(m by a!tial %(ction an# the othe! $o! a gla%% in,ection nee#le to int!o#(ce the 3NA into the male !on(cle(%) The gene!al !oce#(!e $o! mic!oin,ection i% a% $ollow%) 3ono! $emale% a!e in#(ce# to %( e!o"(late (%ing a !o !iate ho!mone t!eatment%) The %( e!o"(late# $emale% a!e then mate# with $e!tile male%' an# la!ge n(mbe!% o$ $e!tiliFe# one/ to two/cell o"aHemb!yo% a!e collecte# %(!gically) Alte!nati"ely' (n$e!tiliFe# o"a a!e collecte# $!om %( e!o"(late# $emale%G the o"a a!e then $e!tiliFe# in "it!o) The t!an% gene con%t!(ct i% !e a!e# in a b($$e! %ol(tion an# i% in,ecte# into the male !on(clei o$ $e!tiliFe# egg% (%ing a mic!oin,ection a%%embly) 6Go!#on et al)-78;9 &" Re$roviral vec$or% Recombinant !et!o"i!(%e% !o#(ce "i!ion%' which a!e (%e# to in$ect

animal cell% an# mice emb!yo%) Ret!o"i!(%e% (%e# a% "ecto!% to t!an%$e! genetic mate!ial into the ho%t cell' !e%(lting in a chime!a' an o!gani%m con%i%ting o$ ti%%(e% o! a!t% o$ #i"e!%e

11

genetic con%tit(tion) Chime!a% a!e inb!e# $o! a% many a% .; gene!ation% (ntil homoFygo(% 6ca!!ying the #e%i!e# t!an%gene in e"e!y cell9 t!an%genic o$$% !ing a!e bo!n) The metho# wa% %(cce%%$(lly (%e# in -7:1 when a %imian "i!(% wa% in%e!te# into mice emb!yo%' !e%(lting in mice ca!!ying thi% 3NA) 6Jaeni%ch et al)-7:19 '" Em(r)onic $em cell Thi% metho# in"ol"e% !io! in%e!tion o$ the #e%i!e# 3NA %e+(ence by homologo(%

!ecombination into an in "it!o c(lt(!e o$ emb!yonic %tem 6ES9 cell%) Stem cell% a!e (n#i$$e!entiate# cell% that ha"e the otential to #i$$e!entiate into any ty e o$ cell 6%omatic an# ge!m cell%9 an# the!e$o!e to gi"e !i%e to a com lete o!gani%m) The%e cell% a!e then inco! o!ate# into an emb!yo at the bla%tocy%t %tage o$ #e"elo ment) The !e%(lt i% a chime!ic animal) ES cell/me#iate# gene t!an%$e! i% the metho# o$ choice $o! gene inacti"ation' the %o/ calle# &noc&/o(t metho#) Thi% techni+(e i% o$ #e"elo mental a!tic(la! im o!tance $o! the %t(#y o$ the genetic cont!ol o$ a!tic(la!ly well in mice) It ha% the a#"antage o$

!oce%%e% an# wo!&%

allowing !eci%e ta!geting o$ #e$ine# m(tation% in the gene "ia homologo(% !ecombination)

E*AMPLES
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13

EIAM@LES: !" Tran +enic Ra((i$ %

Rabbit% a!e +(ite !omi%ing $o! gene $a!ming o! molec(la! $a!ming' which aim% at the !o#(ction o$ !eco"e!able +(antitie% o$ !otein% enco#e# by the t!an% gene%) Gene!ally' the t!an%gene% a!e #e%igne# to be e< !e%%e# in mamma!y ti%%(e %o that thei! !otein ha!"e%te# !o#(ct% a!e %ec!ete# in mil&) Con%e+(ently' the with mil& $!om which it i% !otein !o#(ct i% ea%ily (!i$ie#) con"eniently ha!mace(tically o! biologically im o!tant

The $ollowing h(man gene% enco#ing "al(able

!otein% ha"e been t!an%$e!!e# into

!abbit%: inte!le(&in .' g!owth ho!mone' ti%%(e la%minogen acti"ato!' J-antit!y %in' etc) The%e gene% we!e e< !e%%e# in the mammo!y ti%%(e% an# thei! !otein% we!e ha!"e%te# $!om mil&)

.) Tran +enic Ca$$le:

The only %(cce%%$(l t!an%$ection techni+(e in cattle i% mic!oin,ection o$ $e!tiliFe# o"a' which may eithe! be !eco"e!e# %(!gically o! may be obtaine# $!om o"a!ie% e<t!acte# $!om %la(ghte!e# cow% an# c(lt(!e# in "it!o) The two chie$ ob,ecti"e% o$ t!an%genic !o#(ction a!e a% $ollow%:

6-9 Inc!ea%e# mil& o! meat !o#(ction an# 6.9 molec(la! $a!ming) 609 Se"e!al h(man gene% ha"e been %(cce%%$(lly t!an%$e!!e# in cow% an# e< !e%%e# in the mamma!y ti%%(eG the !otein i% %ec!ete# in mil& $!om whe!e it i% ea%ily ha!"e%te#) '" Tran +enic Pi+
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T!an%genic ig% ha"e al%o been !o#(ce# by $e!tiliFing no!mal egg% with % e!m cell% that ha"e inco! o!ate# $o!eign 3NA) Thi% !oce#(!e' calle# % e!m/me#iate# gene t!an%$e! 6SMGT9 may %ome#ay be able to !o#(ce t!an%genic ig% that can %e!"e a% a %o(!ce o$ t!an% lante# o!gan% $o! h(man%)

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APPLICATIONS

16

17

APPLICATIONS%
1. S$ud)in+ +ene ,unc$ion%

T!an%genic e< !e%%ion i% not only (%e# to b!ee# animal% which ha"e % eci$ic t!ait% b(t al%o to b!ee# animal% that a!e lac&ing % eci$ic gene%) Thi% techni+(e i% calle# a% Knoc&o(t o! Gene ta!geting) T!an%genic technology wa% e!$ecte# (%ing mice in the ea!ly -78;%)Knoc&o(t mice which lac& $(nctional $o!m% o$ % eci$ic gene% a!e !o#(ce# which !o"i#e in$o!mation abo(t the $(nction o$ the &noc&e# o(t gene) Thi% techni+(e allow% %cienti%t% to ta!get % eci$ic gene% $o! the inacti"ation L m(tagene%i%) 6Ko&a!e .;;:9
2. A+ricul$ural A--lica$ion %

a. /reedin+%

5a!me!% ha"e alway% (%e# %electi"e b!ee#ing to !o#(ce animal% that e<hibit #e%i!e# t!ait% 6e)g)' inc!ea%e# mil& !o#(ction' high g!owth !ate9) T!a#itional b!ee#ing i% a time/con%(ming' #i$$ic(lt ta%&) =hen technology (%ing molec(la! biology wa% #e"elo e#' it became o%%ible to #e"elo t!ait% in animal% in a %ho!te! time an# with mo!e !eci%ion) In a##ition' it o$$e!% the $a!me! an ea%y way to inc!ea%e yiel#%) 6Lee et al)-77:9 6Re<!oa# et al)-7879

(. 0uali$)%

T!an%genic cow% e<i%t that !o#(ce mo!e mil& o! mil& with le%% lacto%e o! chole%te!ol-.' ig% an# cattle that ha"e mo!e meat on them an# %hee that g!ow mo!e wool-8) In the a%t' $a!me!% (%e# g!owth ho!mone% to % (! the #e"elo ment o$ animal% b(t thi% techni+(e wa%

18

!oblematic' e% ecially %ince !e%i#(e o$ the ho!mone% !emaine# in the animal !o#(ct) 6ViFe et al)-78:9 6Camme! et al)-7829 c. Di ea e re i $ance% Scienti%t% a!e attem ting to !o#(ce #i%ea%e/!e%i%tant animal%' %(ch a% in$l(enFa/!e%i%tant ig%' b(t a "e!y limite# n(mbe! o$ gene% a!e c(!!ently &nown to be !e% on%ible $o! !e%i%tance to #i%ea%e% in $a!m animal%) 6T!an%genic Li"e%toc& .;;-9 '" Pharmaceu$ical -roduc$ion% 5o! #eca#e%' lowly t!an%genic bacte!ia ha"e %e!"e# a% $acto!ie% $o! many %ca!ce h(man !o#(ct% %(ch a% in%(lin $o! #iabetic% an# g!owth ho!mone% $o! chil#!en who%e bo#ie% #i#n>t ma&e eno(gh) Altho(gh %igni$icantly le%% co%tly than ha!"e%ting the %(b%tance% $!om ti%%(e%' the%e bacte!ial Abio!eacto!%B %till in"ol"e la!ge an# e< en%i"e e+(i ment with com licate# cont!ol%) Othe! %(b%tance% 6e)g)' e!yth!o oietin $o! anemia' bloo# clotting $acto!% $o! hemo hilia an# t@A to t!eat bloo# clot%9 !e+(i!e mo#i$ication% that only cell% o$ highe! o!gani%m% li&e mammal% can !o"i#e) In the a%t' ha!mace(tical com anie% !o#(ce# the%e #!(g% by g!owing mammalian cell% in ti%%(e c(lt(!e) Cowe"e!' thi% i% a "e!y e< en%i"e way to ha!"e%t %mall +(antitie% o$ a %(b%tance) Mil&/ !o#(cing t!an%genic animal% a!e e% ecially (%e$(l $o! !o#(ction o$ me#icine%' n(t! itional %( lement% an# ha!mace(tical%) @!o#(ct% %(ch a% in%(lin' g!owth ho!mone' an# bloo# anti/clotting $acto!% ha"e al!ea#y been obtaine# $!om the mil& o$ t!an%genic cow%' %hee ' o! goat%) Re%ea!ch i% al%o (n#e!way to man($act(!e mil& th!o(gh t!an%gene%i% $o! t!eatment o$ #ebilitating #i%ea%e% %(ch a% henyl&eton(!ia 6@K49' he!e#ita!y em hy%ema' an# cy%tic $ib!o%i%) 6Vete!ina!y =o!l# .;;89 Scienti%t% can now (%e t!an%genic animal% to !o#(ce the%e %(b%tance%) 3(bbe# A ha!ming'B the techni+(e i% $a%t becoming a %igni$icant an# economic alte!nati"e to mo!e con"entional !o#(ction metho#%) Int!o#(cing the gene into a la!ge $a!m animal ha% many a#"antage%: M It i% !e !o#(cible)

19

M The amo(nt o$ #!(g !o#(ce# i% limite# only by the n(mbe! o$ %(cce%%$(l t!an%genic animal% !o#(ce# an# it% o$$% !ing) M Maintaining the animal i% mo!e economical' com a!e# to the amo(nt o$ #!(g !o#(ce#' than g!owing cell% in ti%%(e c(lt(!e)6Boy# et al)-7709 6Camme! et al)-7829 6Me!cie! -78:9

The!e a!e two a !oache% to gene!ating bio!eacto!%' he!e #e$ine# a% t!an%genic animal% !o#(cing ha!mace(tical !otein%) The mo%t e$$ecti"e a !oach i% to e< !e%% a !otein in the mamma!y glan# by (%ing a !omote! $!om a mil& !otein gene to #i!ect e< !e%%ion) An e<am le i% the !o#(ction o$ the o"ine N/lactoglob(lin !omote! $o! (%e in e< !e%%ing a "a!iety o$ h(man !otein% with ha!mace(tical a lication%' %(ch a% J/-/ antit!y %in 6AAT9) Th(%' e< !e%%ion i% #i!ecte# to the mamma!y glan# o$ the lactating mammal' commonly a %hee ' cow' goat o! ig' an# the h(man !otein i% %ec!ete# #i!ectly into the mil&) In the %econ# a !oach' the #e%i!e# the!a e(tic !otein i% !o#(ce# in non/mamma!y bo#y $l(i#%' %(ch a% bloo#) To #ate' thi% a !oach ha% only been (%e# in ig% $o! !o#(cing hemoglobin) 6=all et al)-77E9 6Sha!ma et al)-7719)
4. Indu $rial A--lica$ion %

In .;;-' two %cienti%t% at Ne<ia Biotechnologie% in Cana#a % lice# % i#e! gene% into the cell% o$ lactating goat%) The goat% began to man($act(!e %il& along with thei! mil& an# %ec!ete tiny %il& %t!an#% $!om thei! bo#y by the b(c&et$(l) By e<t!acting olyme! %t!an#% $!om the mil& an# wea"ing them into th!ea#' the %cienti%t% can c!eate a light' to(gh' $le<ible mate!ial that co(l# be (%e# in %(ch a lication% a% milita!y (ni$o!m%' me#ical mic!o%(t(!e%' an# tenni% !ac&et %t!ing%) To<icity/%en%iti"e t!an%genic animal% ha"e been !o#(ce# $o! chemical %a$ety te%ting) Mic!oo!gani%m% ha"e been enginee!e# to !o#(ce a wi#e "a!iety o$ !otein%' which in t(!n can !o#(ce enFyme% that can % ee# ( in#(%t!ial chemical !eaction%) 6B!em et al).;;-9 1" Medical A--lica$ion % a. A a ource o, or+an ,or 2eno$ran -lan$a$ion%
20

@atient% #ie e"e!y yea! $o! lac& o$ a !e lacement hea!t' li"e!' o! &i#ney) 5o! e<am le' abo(t 2';;; o!gan% a!e nee#e# each yea! in the 4nite# King#om alone) T!an%genic ig% may !o"i#e the t!an% lant o!gan% nee#e# to alle"iate the %ho!t$all) C(!!ently' <enot!an% lantation i% ham e!e# by a ig !otein that can ca(%e #ono! !e,ection b(t !e%ea!ch i% (n#e!way to !emo"e the ig !otein an# !e lace it with a h(man !otein) 6Coaglan# et al)-77:9

(. 3uman +ene $hera-)% C(man gene the!a y in"ol"e% a##ing a no!mal co y o$ a gene 6t!an%gene9 to the genome o$ a e!%on ca!!ying #e$ecti"e co ie% o$ the gene) The otential $o! t!eatment% $o! the 2';;; name# genetic #i%ea%e% i% h(ge an# t!an%genic animal% co(l# lay a !ole)

RE4ERENCES

21

RE5ERENCES: -) Be#ell' M)A)' La!gae% a#a' 3)A)' Jen&in%' N)A) L Co elan#' N)G) 6-77:9) Mo(%e mo#el% o$ h(man #i%ea%e) II) Recent !og!e%% an# $(t(!e #i!ection%) Gene% an# 3e"elo ment !!' --)10) .) Boy#' A)L) an# 3) Sami#) 6-7709: Jo(!nal o$ Animal Science : 6%( l) 09: -/7)

0) B!em et al) 6.;;-9 =agne! T!an%genic Li"e%toc&' abo(t t!an%genic metho#%' acce%%e#) 1) B!in%te!' R) 6-7:19) The e$$ect o$ cell% t!an%$e!!e# into mo(%e bla%tocy%t on %(b%e+(ent #e"elo ment) J) E< ) Me#):-;17/-;2E) 2) Echela!# ?) 6-77E9) Recombinant O inion in Biotechnology :: 20E/21;) E) 5ELASA 6-78.9 5e#e!ation o$ E(!o ean Labo!ato!y Animal Science A%%ociation%) T!an%genic Animal% 3e!i"ation' =el$a!e' 4%e an# @!otection) :) 5(lton S) 6.;;;9) Ro(n#( on bio !oce%% "ali#ation i%%(e%: t!an%genic animal !o#(ction o$ bio ha!mace(tical%) Genetic Enginee!ing New%' ) .;:0E !otein !o#(ction in t!an%genic animal%) C(!!ent

22

8) Go!#on' J)=) an# R(##le' 5)C) 6-78-9) Integ!ation an# %table ge!m line t!an%$o!mation o$ gene% in,ecte# into mo(%e !on(clei) Science &!5:-.11/-.1E) 7) Go%%le!' A) et al) 6-78E9) T!an%gene%i% by mean% o$ bla%tocy%t/#e!i"e# emb!yonic %tem cell line) @!oc) Natl) Aca#) Sci) 6':7;E2/7;E7)
10. Camme! R)E)' V)G) @(!%el' C)E) Re<!oa#' R)J) =all' 3)J) Bolt' K)M) Ebe!t' R)3)

@almite! an# R)L) B!in%te!) 6-7829) A@!o#(ction o$ t!an%genic !abbit%' %hee ' an# ig% by mic!oin,ection)B Nat(!e 0-2: E8;/E80
11. Coaglan#' T)A)' M) J(lian' J)=) Rie%en' 3) Sch!iebe! an# =)L) 5o#o! 6-77:9)

AT!an%genic ig% a% animal mo#el $o! <enogenic t!an% lantation)B The!iogenology 1:: ..1 6Ab%t!act9 -.) Jaeni%ch' R) 6-7:E9) Ge!m line integ!ation an# Men#elian t!an%mi%%ion o$ the e<ogeno(% Moloney le(&emia "i!(%) @!oc) Natl) Aca#) Sci) 7':-.E;/-.E1)
13. Lee' C)S)' ?)C) Choi' K)B) Oh' ?)K) Kang an# K)K) Lee) 6-77:9) ATem o!al an# % atial

% eci$ic e< !e%%ion o$ bo"ine b/ca%einHbo"ine g!owth ho!mone $(%ion gene in t!an%genic mice)B The!iogenology 1:: .2 6Ab%t!act9) -1) Me!cie!' J)C) 6-78:9: AGenetic enginee!ing a lie# to mil&/ !o#(cing animal%: %ome e< ectation%)B E< loiting New Technologie% in Animal B!ee#ing' ) -../-0-) -2) Ko&a!e C)R) 6.;;:9) @ha!mace(tical BiotechnologyG 5(n#amental% an# A lication%G Ni!ali !a&a%hanG -0)-./-0)-E) -E) @hili Le#e!: Vignette% 6.;;09 La%&e! 5o(n#ation>% bio abo(t Le#e!' lea# %cienti%t $o! OncoMo(%e)
17. @(!%el V)G) ' Re<!oa# C)E' J!)' Bolt 3)J' K)5) Mille!' )J) =all' R)E) Camme!' C)A)

@in&e!t' )3) @almite! an# )L) B!in%te!) 6-78:9) A@!og!e%% on gene t!an%$e! in $a!m animal%)B Vete!ina!y Imm(nology Imm(no athology -:: 0;0/0-.

23

-8) Reggio' B)C)' C)L) G!een' M) San%inena' L)C) Chen' E) Behboo#i' R)S) 3enni%ton' ?) Echela!# an# R)A) Go#&e) 6.;;.9) @!o#(ction o$ clone# t!an%genic goat% a% a otential %o(!ce $o! h(man ha!mace(tical%) The!iogenology 2::112)
19. Re<!oa#' C)E)' J!)' R)E) Came!' 3)J) Bolt' K)E) Mayo' A) 5!ohman' R)3) @amite! an#

R)L) B!in%te!) 6-7879) A@!o#(ction o$ t!an%genic %hee with g!owth/!eg(lating gene%)B Molec(la! Re !o#(cti"e 3e"elo ment -: -E1 6Ab%t!act9
20. Re<!oa#' C)E)' J!)' R)E) Came!' 3)J) Bolt' K)E) Mayo' A) 5!ohman' R)3) @amite! an#

R)L) B!in%te!) 6-7879 A@!o#(ction o$ t!an%genic %hee with g!owth/!eg(lating gene%)B Molec(la! Re !o#(cti"e 3e"elo ment -: -E1 6Ab%t!act9
21. Ro%chla(' K)' @) Rommel' L) An#!eewa' M) Oac&el' 3) Ro%chla(' B) Oac&el' M)

Schwe!in' M) C(hn an# K)G) GaFa!,an) 6-7879) AGene t!an%$e! e< e!iment% in cattle)B Jo(!nal o$ Re !o#(ction an# 5e!tility 6S( l)9 08: -20/-E;

..) Scott Gottlieb 6.;;.9) Con%e+(ence% o$ the Biotechnology Re"ol(tion' New ?o!& S(n' A--) .0) Sha!ma' A)' Ma!tin' M)J)' O&abe' J)5)' T!(glio' R)A)' 3han,al' N)K)' Logan' J)S) L Kh(ma!' R) 6-7719) An i%ologo(% o!cine !omote! e!mit% high le"el e< !e%%ion o$ h(man hemoglobin in t!an%genic %wine) Biotechnology !&' 22)27) .1) t!an%genic animal% an# ta!gete# m(tation%) Nat(!e '8'' 0:2)0:E) .2) T!an%genic Animal% 6.;;09' 5(n# $o! the Re lacement o$ Animal% in Me#ical E< e!iment%' 5RAME' $act %heet abo(t t!an%gene%i% an# i%%(e%) .E) T!an%genic Li"e%toc& 6.;;-9' (n#ate# a!ticle by G) B!em an# C)G) =agne! abo(t t!an%genic metho#%) .:) Vete!ina!y =o!l# 6.;;89 Vol)- 6-;9: 0-7/0.; .8) ViFe' @)3)' A) Michal%&a' R) A%hman' R)5) Seama!& an# J)R)E) =ell%) 6-78:9 AIm !o"ing g!owth in t!an%genic $a!m animal%)B EMBO =o!&%ho Mani (lation o$ Animal%) Nethyb!i#ge' Scotlan#' 4K) .7) =all' R)J) 6-77E9) Mo#i$ication o$ mil& com o%i/ tion in t!an%genic animal%) In Biotechnology)% Role in the Genetic Im !o"ement o$ 5a!m Animal% 6e#) R)C) Mille!' on Ge!mline

24

V)G) @(!%ell L C)3) No!man9' Animal Science)

) -E2)-88) Sa"oy' IL' 4SA: The Ame!ican Soci/ ety o$

0;) =oychic&' R)@)' =a%%om' J)S) L King%b(!y' 3) 6-7709) TBASE: a com (te!iFe# #ataba%e $o! 0-) ?o(ng' M)=)' C) Mea#e' J) C(!ling' C) Oiome&' an# M) Ca!"ey) 6-7789) @!o#(ction o$ !ecombinant antibo#ie% in the mil& o$ t!an%genic animal%) Re% Imm(nol) J(l/A(gG -176E9: E;7/E-;)

25

TRANSGENIC @LANTS

26

OBJECTIVE

OBJECTIVE: -) The aim o$ c!eating t!an%genic lant% i% to #e%ign lant% with % eci$ic cha!acte!i%tic% th!o(gh a!ti$icial in%e!tion o$ gene% $!om othe! % ecie%)

27

2. T!an%genic

lant% ha"e been enginee!e# to o%%e%% %e"e!al #e%i!able t!ait%' incl(#ing e%t%' he!bici#e% o! ha!%h en"i!onmental con#ition%' im !o"e# !o#(ct

!e%i%tance to

%hel$li$e' an# inc!ea%e# n(t!itional "al(e) 0) The !o#(ction o$ $o!eign !otein% in lant% ha% become an att!acti"e alte!nati"e to con"entional !o#(ction %y%tem% $o! ha!mace(tical oly e ti#e%) 1) A !e#(ction in the o"e!all amo(nt o$ e%tici#e% (%e# in c!o a %a$e! en"i!onment) !o#(ction' which im act%

o%iti"ely on bio#i"e!%ity' !otect% !e#ato!% an# non/ta!get o!gani%m% an# cont!ib(te% to

28

INTRO34CTION

INTRO34CTION:

29

The!e wa% a time not too long ago when mo%t me#icinal com o(n#% came $!om lant%: the otent hea!t %tim(lant #igitali% $!om $o<glo"e' $o! e<am le' an# o i(m $!om o y lant) B(t beginning abo(t 2; yea!% ago' chemi%t!y too& o"e! $!om botany' with mo%t new #!(g% being a!ti$icially !o#(ce# in ha!mace(tical lab%) Nowa#ay%' one o$ the mo%t !omi%ing metho#% o$ !o#(cing !otein% an# othe! me#icinal %(b%tance%' %(ch a% antibo#ie% an# "accine%' i% the (%e o$ t!an%genic lant%) 6Mo$$at -7729 Tran +enic -lan$ a!e lant% o%%e%%ing a %ingle o! m(lti le gene%' t!an%$e!!e# $!om a #i$$e!ent % ecie%) Tho(gh 3NA $!om anothe! % ecie% can be integ!ate# into a lant*% genome "ia nat(!al !oce%%e%' the te!m Pt!an%genic lant%P !e$e!% to lant% c!eate# in a labo!ato!y (%ing !ecombinant 3NA technology) 65e%chotte et al).;;29 The aim o$ c!eating t!an%genic lant% i% to #e%ign lant% with % eci$ic cha!acte!i%tic% th!o(gh a!ti$icial in%e!tion o$ gene% $!om othe! % ecie% 6o! ta<onomically ( to #i$$e!ent &ing#om%9) A t!an%genic lant contain% a gene o! gene% that ha"e been a!ti$icially in%e!te#) The in%e!te# gene %e+(ence' &nown a% the t!an%gene' may come $!om an (n!elate# lant' o! $!om a com letely #i$$e!ent % ecie%) One o$ the (! o%e% o$ in%e!ting a combination o$ gene% in a lant i% to ma&e it a% (%e$(l an# !o#(cti"e a% o%%ible) Thi% !oce%% !o"i#e% a#"antage% %(ch a% highe! yiel#' im !o"e# +(ality' e%t o! #i%ea%e !e%i%tance' an# tole!ance to heat' col# an# #!o(ght) Cowe"e!' t!an%genic lant% can al%o be !o#(ce# in %(ch a way that they e< !e%% $o!eign !otein% with in#(%t!ial o! ha!mace(tical "al(e) T!an%genic lant% !e !e%ent an economical alte!nati"e to $e!mentation/ba%e# !o#(ction %y%tem%) @lant/ma#e "accine% o! antibo#ie% 6 lantibo#ie%9 a!e e% ecially %t!i&ing' a% lant% a!e $!ee o$ h(man #i%ea%e%' th(% !e#(cing %c!eening co%t% $o! "i!(%e% an# bacte!ial to<in%) 6Ce!be!% et al )-7779) The $i!%t t!an%genic lant% we!e !e o!te# in -780) Since then' many !ecombinant !otein% ha"e been e< !e%%e# in %e"e!al im o!tant ag!onomic % ecie% o$ lant% incl(#ing tobacco' co!n' tomato' otato' banana' al$al$a an# canola) The choice o$ we!e gene!ally (%e#' howe"e!' lant %y%tem wa% initially #!i"en by (! o%e o$ con"enience an# the nee# to e"al(ate genetic con%t!(ct% +(ic&ly) 5o! thi% !ea%on' tobacco lant% otatoe% an# banana% we!e con%i#e!e# $o! the ac+(i!ing a #eli"e!y %y%tem o$ "accine% $o! h(man%) 6Cammon# et al) -7779 5o!eign gene e< !e%%ion in lant% can be accom li%he# eithe! by %table integ!ation o$ $o!eign
30

3NA' which !e%(lt% in t!an%$o!mation o$ the n(clea! genome' o! by t!an%ient e< !e%%ion (%ing mo#i$ie# lant "i!(%e%) Stable genomic integ!ation i% accom li%he# by int!o#(cing $o!eign 3NA in the lant eithe! by Ag!o bacte!i(m T/3NA "ecto!% o! by #i!ect mean%) The integ!ation metho# ha% the a#"antage o$ e!mitting la!ge %cale cloning' maintenance o$ %electe# high/e< !e%%ing gene% an# the ability to %e<(ally c!o%% t!an%gene to obtain m(lti le !otein% e< !e%%e# in the %ame lant) On the othe! han#' t!an%ient e< !e%%ion (%ing "i!al "ecto!% i% ha!#e! to initiate' beca(%e the "i!al "ecto! m(%t be inoc(late# into in#i"i#(al ho%t lant%' b(t gi"e% a g!eate! yiel# o$ !otein) 6Tac&et et al) -7789

31

TRANSGENIC @ROCESS

32

TRANSGENIC @ROCESS:
T!an%genic lant% can be !o#(ce# by two metho#%: -) Vecto! me#iate# gene t!an%$e! 6a9 6b9 .) Bacte!ial "ecto! Vi!(% "ecto!

3i!ect gene t!an%$e! 6a9 Elect!o o!ation 6b9 Li o%ome me#iate# gene t!an%$e! 6c9 Mic!oin,ection 6#9 Mac!oin,ection

33

6e9 Balli%tic% OR @a!ticle G(n $o! Gene T!an%$e! 6$9 @ollen OR @ollen T(be 6g9 4lt!a%onication !"Vec$or media$ed +ene $ran ,er @lant% #e"elo c!own gall% ( on in$ection with Ag!obacte!i(m' an# thi% in"ol"e% the t!an%$e! o$ gene% $!om the bacte!i(m to the lant) Ag!obacte!i(m t(me$acien% i% a ty e o$ bacte!ia that t!an%$e!% it% 3NA "ia ho!iFontal gene t!an%$e! to c!eate t(mo!% in ag!obacte!i(m ha lant%) Thi% ma&e% it "e!y (%e$(l to genetic enginee!ing) Gene t!an%$e! (%ing en% when Aa !e%t!iction enFyme i% (%e# to c(t non/"i!(lent la%mi# 3NA #e!i"e#

$!om A) t(me$acien% an# th(% c!eate an in%e!tion oint' into which the gene can be ligate#) The enginee!e# la%mi# i% then (t into a %t!ain o$ A) t(me$acien%' which contain% a Qhel e!> la%mi# an# lant cell% a!e t!eate# with the !ecombinant bacte!i(mB in c(lt(!e) R2S =hile thi% loo&% li&e a com licate# conce t' it>% !eally only a genetic enginee!ing "e!%ion o$ c(t an# a%te) 63omingo .;;:9

&"

Direc$ +ene $ran ,er 9a. Elec$ro-ora$ion% Thi% metho# i% ba%e# on the (%e o$ %ho!t elect!ical im (l%e% o$ high $iel# %t!ength) The%e im (l%e% inc!ea%e the e!meability o$ !oto la%t memb!ane an# $acilitate% ent!y o$ 3NA molec(le% into the cell%' i$ the 3NA i% in #i!ect contact with the memb!ane) In "iew o$ thi%' $o! #eli"e!y o$ 3NA to !oto la%t%' elect!o o!ation i% one o$ the %e"e!al !o(tine techni+(e% $o! e$$icient t!an%$o!mation) The elect!o o!ation (l%e i% gene!ate# by #i%cha!ging a ca acito! ac!o%% the elect!o#e% in a % ecially #e%igne# elect!o o!ation chambe!) Eithe! a high "oltage 6-)2 &V9 !ectang(la! wa"e (l%e o$ %ho!t #(!ation o! a low "oltage 602;V9 (l%e o$ long #(!ation i% (%e#) @!oto la%t% in an ionic %ol(tion containing the "ecto! 3NA' a!e %(% en#e# between the elect!o#e%' elect!o o!ate# an# then late# a% (%(al) 4%ing elect!o o!ation metho#' %(cce%%$(l t!an%$e! o$ gene% wa% achie"e# with the !oto la%t% o$ tobacco' et(nia' maiFe' !ice' wheat an# %o!gh(m) In mo%t o$ the%e ca%e% cat gene a%%ociate# with a %(itable !omote! %e+(ence wa% t!an%$e!!e#)

34

T!an%$o!mation $!e+(encie% can be $(!the! im !o"e# by 6i9 4%ing $iel# %t!ength o$ -).2 &VHcm 6ii9 A##ing @EG a$te! a##ing 3NA 6iii9 Ceat %hoc&ing !oto la%t% at 12TC $o! 2 min(te% be$o!e a##ing 3NA 6i"9 By (%ing linea! in%tea# o$ ci!c(la! 3NA 6Chan# et al)-7889)

9(. Li-o ome media$ed +ene $ran ,er% Li o%ome% a!e %mall li i# bag%' in which la!ge n(mbe! o$ la%mi#% a!e enclo%e#) They can be in#(ce# to $(%e with !oto la%t% (%ing #e"ice% li&e @EG' an# the!e$o!e ha"e been (%e# $o! gene t!an%$e!) The techni+(e' o$$e!% $ollowing a#"antage%: 6i9 @!otection o$ 3NAIRNA $!om n(clea%e #ige%tion' 6ii9 Low cell to<icity' 6iii9 Stability an# %to!age o$ n(cleic aci#% #(e to enca %(lation in li o%ome%' 6i"9 Cigh #eg!ee o$ !e !o#(cibility an# 6"9 A licability to a wi#e !ange o$ cell ty e%)

In thi% techni+(e' 3NA ente!% the $ollowing %te %:

!oto la%t% #(e to en#ocyto%i% o$ li o%ome%' in"ol"ing the

6i9 a#he%ion o$ the li o%ome% to the !oto la%t %(!$ace 6ii9 $(%ion o$ li o%ome% at the %ite o$ a#he%ion 6iii9 !elea%e o$ la%mi#% in%i#e the cell) The techni+(e ha% been %(cce%%$(lly (%e# to #eli"e! 3NA into the !oto la%t% o$ a n(mbe! o$ lant % ecie% 6e)g) tobacco' et(nia' ca!!ot' etc)9)

6c9

Microin#ec$ion% =hen cell% o!

!oto la%t% a!e (%e# a% ta!get% in the techni+(e o$ a!e (%e# $o! t!an%$e! o$

mic!oin,ection' gla%% mic!o i ette% with ;)2/-;Um #iamete! ti a!e immobiliFe# on a %oli# %(

mac!omolec(le% into the cyto la%m o! the n(cle(% o$ a !eci ient cell o! !oto la%t) The !eci ient cell% o!t 6co"e! %li o! %li#e9 o! a!ti$icially bo(n# to a %(b%t!ate o! hel# by a

35

i ette (n#e! %(ction 6a% #one in animal %y%tem%9)O$ten a % ecially #e%igne# mic!omani (lato! i% em loye# $o! mic!oin,ecting the 3NA) Altho(gh' thi% techni+(e gi"e% high !ate o$ %(cce%%' the !oce%% i% %low' e< en%i"e an# !e+(i!e% highly %&ille# an# e< e!ience# e!%onnel)

6#9

Macroin#ec$ion% 3NA mac!oin,ection em loying nee#le% with #iamete!% g!eate! than cell

#iamete! ha% al%o been t!ie#) In !ye' a ma!&e! gene wa% mac!oin,ecte# into the %tem below the immat(!e $lo!al me!i%tem' %o a% to !each the % o!ogeno(% ti%%(e lea#ing to %(cce%%$(l !o#(ction o$ t!an%genic lant%) 6e9 /ioli $ic OR Par$icle Gun ,or Gene Tran ,er% / In !ecent yea!%' it ha% been %hown that 3NA #eli"e!y to lant cell% i% al%o o%%ible' when hea"y mic!o a!ticle% 6t(ng%ten o! gol#9 coate# with the 3NA o$ inte!e%t a!e accele!ate# to a "e!y high initial "elocity) Thi% i% %ometime% calle# Pbioli%tic%P / a c!o%% between biology an# balli%tic%) Anothe! te!m $o! the #e"ice i% a Pgene g(nP

36

The%e mic!o !o,ectile%' no!mally -/0 m in #iamete!' a!e ca!!ie# by a *mac!o !o,ectile* o! the *b(llet* an# a!e accele!ate# into li"ing lant cell% 6ta!get cell% can be ollen' c(lt(!e# cell%' cell% in #i$$e!entiate# ti%%(e% an# me!i%tem%9 %o that they can enet!ate cell wall% o$ intact ti%%(e) T!an%$o!me# lant% (%ing the abo"e techni+(e ha"e been obtaine# in many ca%e% incl(#ing %oybean' tobacco' maiFe' !ice' wheat' etc)) T!an%ient e< !e%%ion o$ gene% t!an%$e!!e# in cell% by thi% metho# ha% al%o been ob%e!"e# in onion' maiFe' !ice an# wheat)

(f) t(be

Pollen OR -ollen Tu(e% The!e ha% been a ho e that 3NA can be ta&en ( by the athway) Both the%e a !oache% ha"e been t!ie# an# inte!e%ting henoty ic alte!ation%

ge!minating ollen an# can eithe! integ!ate into % e!m n(clei o! !each the Fygote th!o(gh the ollen %(gge%ting gene t!an%$e! ha"e been obtaine#) In no ca%e' howe"e!' (ne+(i"ocal !oo$ o$ gene t!an%$e! ha% been a"ailable) In a n(mbe! o$ e< e!iment%' when ma!&e! gene% we!e (%e# $o! t!an%$e!' only negati"e !e%(lt% we!e obtaine#) Se"e!al !oblem% e<i%t in thi% metho# an# the%e incl(#e the !e%ence o$ cell wall' n(clea%e%' hete!och!omatic %tate o$ acce to! 3NA' callo%e l(g% in ollen t(be' etc) 6Ci%to!y o$ @lant B!ee#ing .;;8.

37

6g9

:l$ra onica$ion% In wheat' tobacco an# %(ga!beet' e< lant% a$te! being c(lt(!e# $o! a $ew

#ay%' we!e %onicate# with la%mi# 3NA which ca!!ying ma!&e! gene%) =hen %onicate# calli we!e t!an%$e!!e# to %electi"e me#i(m' %hoot% we!e obtaine#' altho(gh all cont!ol calli 6not %onicate# with la%mi# 3NA9 #ie#) In tobacco t!an%genic lant% we!e obtaine# at a $!e+(ency o$ .. e! cent) Thi% wo!& wa% con#(cte# at the Biotechnology Re%ea!ch Cente! at Bei,ing 6China9) 6Sta#le! an# S !ag(e -70E9 6Be!g et al).;;0.

38

E*AMPLES

39

EIAM@LES: !" Tran +enic rice%

Tran +enic rice i% a ty e o$ !ice that ha% (n#e!gone genetic mo#i$ication% $o! ag!ic(lt(!al (! o%e%) Nat(!al mo"ement o$ gene% between % ecie%' o$ten calle# ho!iFontal gene t!an%$e! o! late!al gene t!an%$e!' can occ(! with !ice beca(%e o$ gene t!an%$e! me#iate# by nat(!al !oce%%e%) The!e i% new %cienti$ic lite!at(!e abo(t nat(!al t!an%genic e"ent% in lant%' th!o(gh mo"ement o$ nat(!al mobile 3NA% calle# M4LE% between !ice an# Seta!ia millet) Gol#en !ice i% a "a!iety o$ O!yFa %ati"a !ice !o#(ce# th!o(gh genetic enginee!ing to

bio%ynthe%iFe beta/ca!otene' a !ec(!%o! o$ !o/"itamin A in the e#ible a!t% o$ !ice) The %cienti$ic #etail% o$ the !ice we!e $i!%t (bli%he# in Science in .;;;) New Rice $o! A$!ica 6PNERICAP9 i% an inte!% eci$ic c(lti"a! o$ !ice #e"elo e# by the =e%t A$!ica Rice 3e"elo ment A%%ociation 6=AR3A9 to im !o"e the yiel# o$ A$!ican !ice "a!ietie%) 6@LoS Biology .;;E9

40

&" Tran +enic o)(ean%

The $ran +enic o)(ean' al%o &nown a% the !ecombinant %oybean' genetically enginee!e# %oybean' o! genetically mo#i$ie# %oybean' i% a %oybean 6Glycine ma<9 that ha% ha# 3NA int!o#(ce# into it in a way othe! than the combination o$ a % e!m an# an egg) Thi% !oce%% i% ca!!ie# o(t (%ing A!ecombinant 3NA 6!3NA9 technology)BThi% technology i% not limite# to !o#(cing only the t!an%genic %oybean' b(t to !o#(cing genetically mo#i$ie# o!gani%m% 6GMO%9 o$ any lant' animal' o! mic!obial "a!iety) The %oybean' howe"e!' i% "e!y "e!%atile in it% (%e' th(% ma&ing it a o (la! %(b,ect to genetic mo#i$ication)6Celec et al) .;;29 6Anthony -7789

'" Tran +enic ;hea$%

=heat 6T!itic(m %

)9 i% an im o!tant #ome%ticate# g!a%% (%e# wo!l#wi#e $o! $oo#' an# it%

e"ol(tion ha% been in$l(ence# by h(man inte!"ention %ince the #awn o$ ag!ic(lt(!e) =heat i% a nat(!al hyb!i# #e!i"e# $!om inte!% ecie% b!ee#ing) It i% theo!iFe# that wheat*% ance%to!% 6T!itic(m monococc(m' Aegilo % % eltoi#e%' an# Aegilo % ta(%chii' all #i loi# g!a%%e%9 hyb!i#iFe# nat(!ally o"e! millennia %omewhe!e in =e%t A%ia' to c!eate nat(!al oly loi# hyb!i#%' the be%t &nown o$ which a!e common wheat an# #(!(m wheat) 6=!igley .;;19 5" /$ (rin#al%

/$ (ai+un i% a t!an%genic b!in,al c!eate# by in%e!ting a gene RC!y-AcS $!om the %oil bacte!i(m Bacill(% th(!ingien%i% into b!in,al) The in%e!tion o$ the gene into the b!in,al cell in yo(ng cotyle#on% ha% been #one th!o(gh an Ag!o bacte!i(m/me#iate# "ecto!' along with othe! gene% li&e !omote!%' ma!&e!% etc) Thi% i% %ai# to gi"e the b!in,al lant !e%i%tance again%t le i#o te!on in%ect% li&e the B!in,al 5!(it an# Shoot Bo!e! 6Le(cino#e% o!bonali%9 an# 5!(it Bo!e! 6Celico"e! a a!mige!a9) It i% !e o!te# that ( on inge%tion o$ the Bt to<in by the in%ect'

41

the!e wo(l# be #i%!( tion o$ #ige%ti"e !oce%%e%' (ltimately !e%(lting in the #eath o$ the in%ect) 6B(!che! .;;79)

Mon%anto' an Ame!ican com any' ha% #e"elo e# the Bt b!in,al an# it i% being ma!&ete# in In#ia by MH% Mahyco RMaha!a%ht!a Cyb!i# See#% Com anyS)Now' the com any want% to ta&e ( la!ge %cale $iel# t!ial% with the e!mi%%ion o$ the GEAC in .;;E/;:) The im o!tance o$ thi% #e"elo ment can be (n#e!%too# $!om the $act that no GM b!in,al ha% been !elea%e# $o! an a#"ance# %tage o$ $iel# t!ial% in o en con#ition% anywhe!e in the wo!l# an# that thi% i% the $i!%t time that GEAC co(l# be gi"ing e!mi%%ion $o! la!ge %cale o en t!ial% $o! a $oo# c!o )

42

APPLICATIONS

43

A@@LICATIONS:
T!an%genic lant can be mani (late# to act a% bio!eacto!% to !o#(ce a wi#e !ange o$ biologically' im o!tant com o(n#%) The comme!cial !o#(ct o$ lant% i% (%e$(l $o! in#(%t!ie% o! $o! im !o"ing the h(man o! animal health) @lant genetic enginee!ing ha% ma#e o%%ible the t!an%$e! o$ gene% ac!o%% all ta<onomic ba!!ie!) Gene% $!om bacte!ia' $(ngi' in%ect% an# "i!(%e% ha"e been int!o#(ce# in to lant%) The (ltimate goal o$ t!an%gene i% to im !o"e the c!o %' with #e%i!e# t!ait% li&e im !o"e the c!o yiel#' an# +(ality e)g) %to!age' longe!' %el$ li$e o$ $!(it% an# $lowe!%) T!an%genic lant% o$$e! a new a chea an# ea%y to a#mini%te!) The #i$$e!ent a lication% o$ t!an%genic lant% a!e #e%c!ibe# a% $ollow%: !oach to !o#(cing an# a#mini%te!ing h(man antibo#ie%) The !ecent

!e%ea!ch with t!an%genic lant% ha% laye# a ca ti"ating !ole in !o"i#ing e#ible "accine%' which a!e

!" Molecular -harmin+ Molec(la! $a!ming i% a ty e o$ genetic mo#i$ication (%e# in $a!ming in"ol"ing the (%e o$ lant%' an# otentially al%o animal%' a% the mean% to $a!ming .;;8. The $i!%t !ecombinant lant/#e!i"e# ha!mace(tical !otein 6@3@9 wa% h(man %e!(m alb(min' !o#(ce com o(n#% o$ the!a e(tic "al(e) 6Molec(la!

initially !o#(ce# in -77; in t!an%genic tobacco an# otato lant%) 6Twyman an# Richa!# .;;09) 5i$teen yea!% on' the $i!%t technical !otein% !o#(ce# in t!an%genic lant% a!e on the ma!&et' an# !oo$ o$ conce t ha% been e%tabli%he# $o! the !o#(ction o$ many the!a e(tic !otein%' incl(#ing antibo#ie%' bloo# !o#(ct%' cyto&ine%' g!owth $acto!%' ho!mone%' !ecombinant enFyme% an# h(man an# "ete!ina!y "accine%) 6Ch!i%to( et al).;;09) 5(!the!mo!e' %e"e!al @3@ !o#(ct% $o! the t!eatment o$ h(man #i%ea%e% a!e a !oaching comme!cialiFation' incl(#ing !ecombinant ga%t!ic li a%e $o! the i elineG 3ow t!eatment o$ cy%tic $ib!o%i%' an# antibo#ie% $o! the !e"ention o$ #ental ca!ie% an# the t!eatment o$ non/Co#g&in*% lym homa)The!e a!e al%o %e"e!al "ete!ina!y "accine% in the

44

Ag!oScience% anno(nce# !ecently thei! intention to !o#(ce lant/ba%e# "accine% $o! the animal health in#(%t!y) Global #eman# $o! ha!mace(tical% i% at (n !ece#ente# le"el%' an# c(!!ent !o#(ction ca acity will %oon be o"e!whelme#) E< an#ing the e<i%ting mic!obial %y%tem%' altho(gh $ea%ible $o! %ome the!a e(tic !o#(ct%' i% not a %ati%$acto!y o tion on %e"e!al g!o(n#%) 5i!%t' it wo(l# be "e!y e< en%i"e $o! the ha!mace(tical com anie%) Secon#' othe! !otein% o$ inte!e%t a!e too com le< to be ma#e by mic!obial %y%tem%) The%e !otein% a!e c(!!ently being !o#(ce# in animal cell c(lt(!e%' b(t the !e%(lting !o#(ct i% o$ten !ohibiti"ely e< en%i"e $o! many atient%) 6Ch!i%to( et al).;;09 &" A+ricul$ural u e Ag!ic(lt(!e i% the !o#(ction o$ $oo# an# goo#% th!o(gh $a!ming) The %t(#y o$ ag!ic(lt(!e i% &nown a% ag!ic(lt(!al %cience) The ma,o! ag!ic(lt(!al !o#(ct% can be b!oa#ly g!o( e# into $oo#%' $ibe!%' $(el%' an# !aw mate!ial%) In the .;;;%' lant% ha"e been (%e# to g!ow bio$(el%' bio ha!mace(tical%' bio la%tic%) @la%tic% that a!e g!een in mo!e way% than oneP6=a%hington 3C .;;79 In .;;:' abo(t one thi!# o$ the wo!l#*% wo!&e!% we!e em loye# in ag!ic(lt(!e) The %e!"ice% %ecto! ha% o"e!ta&en ag!ic(lt(!e a% the economic %ecto! em loying the mo%t eo le wo!l#wi#e) 6Key In#icato!% o$ the Labo(! Ma!&et @!og!amme .;;79 9a. 3er(icide<$oleran$ GMO CroRo(n#( /Rea#y %ee# ha% a he!bici#e !e%i%tant gene im lante# into it% genome that allow% the lant% to tole!ate e< o%(!e to gly ho%ate) Ro(n#( i% a t!a#e name $o! a gly ho%ate ba%e# !o#(ct' which i% a %y%temic' non/%electi"e he!bici#e (%e# to &ill wee#%) Ro(n#( /Rea#y %ee#% allow the $a!me! to g!ow a c!o that can be % !aye# with gly ho%ate to cont!ol wee#% witho(t ha!ming the !e%i%tant c!o ) Ce!bici#e/tole!ant c!o % a!e (%e# by $a!me!% wo!l#wi#e) To#ay' 7.V o$ %oybean ac!eage in the 4S i% lante# with genetically/mo#i$ie# he!bici#e/tole!ant lant%) =ith the inc!ea%ing (%e o$ he!bici#e/ tole!ant c!o %' come% an inc!ea%e in the (%e o$ gly ho%ate ba%e# he!bici#e % !ay%) In %ome a!ea% gly ho%ate !e%i%tant wee#% ha"e #e"elo e#' ca(%ing $a!me!% to %witch to othe! he!bici#e) Some %t(#ie% al%o lin& wi#e% !ea# gly ho%ate (%age to i!on #e$iciencie% in %ome c!o %' which i% both a c!o !o#(ction an# a n(t!itional +(ality conce!n' with otential economic an# health im lication%) 6A#o tion o$ Genetically Enginee!e# C!o % in the 4S: E<tent o$ A#o tion .;;89 65a!me!% G(i#e to GMO% .;;89 6Re o!t Rai%e% Ala!m o"e! *S( e!/wee#% .;;89 6OFt(!&' et al).;;89

45

9(. In ec$<Re i $an$ GMO CroOthe! GMO c!o % (tiliFe# by g!owe!% incl(#e in%ect/!e%i%tant c!o %' which ha"e a gene $!om the %oil bacte!i(m Bacill(% th(!ingien%i% 6Bt9' which !o#(ce% a to<in % eci$ic to in%ect%) The%e c!o % !otect lant% $!om #amage by in%ect%G one %(ch c!o i% Sta!lin&) Anothe! i% cotton' which acco(nt% $o! E0V o$ 4S cotton ac!eage) Some belie"e that %imila! o! bette! e%t/!e%i%tance t!ait% can be ac+(i!e# th!o(gh t!a#itional b!ee#ing !actice%' an# !e%i%tance to "a!io(% e%t% can be gaine# th!o(gh hyb!i#iFation o! c!o%%/ ollination with wil# % ecie%) In %ome ca%e%' wil# % ecie% a!e the !ima!y %o(!ce o$ !e%i%tance t!ait%G %ome tomato c(lti"a!% that ha"e gaine# !e%i%tance to at lea%t nineteen #i%ea%e% #i# %o th!o(gh c!o%%ing with wil# o (lation% o$ tomatoe%)

46

6Genetically Enginee!e# C!o % in the 4S .;;89 6Kimb!ell .;;.9

47

RE5ERENCES

48

RE5ERENCES:

1.

A#o tion o$ Genetically Enginee!e# C!o % in the 4S: E<tent o$ A#o tion ) Ret!ie"e# 3ecembe! 8' .;;8)

.)

Anthony' Kinney J) an# S(%an Knowlton 6-7789) A3e%igne! Oil%: The Cigh Oleic Aci# Soybean)B Genetic Mo#i$ication in the 5oo# In#(%t!y: A St!ategy $o! 5oo# W(ality Im !o"ement) E#) Rolle!' Sibel an# S(%an Ca!lan#e!) Lon#on: Blac&ie) -70/.-0)

0) 1)
5.

Be!g' @a(lG Singe!' Ma<ine 6A(g(%t -2' .;;09) Geo!ge Bea#le: An 4ncommon 5a!me!) The Eme!gence o$ Genetic% in the .;th cent(!y) Col# S !ing% Ca!bo! Labo!ato!y @!e%%) Be!g' @a(lG Singe!' Ma<ine 6A(g(%t -2' .;;09) Geo!ge Bea#le: An 4ncommon 5a!me!) The Eme!gence o$ Genetic% in the .;th cent(!y) Col# S !ing% Ca!bo! Labo!ato!y @!e%%) B(!che! 6.;;79) PBt B!in,al 4n$it $o! C(man Con%(m tion / E< e!t in#e en#ent %cienti%t "e!#ict cont!a#ict% In#ian !eg(lato!y a(tho!itie% on bio%a$ety o$ GM a(be!gineP) htt :HHwww)i/%i%)o!g)(&HBtXB!in,alX4n$it) h )

E) Celec et al) 6.;;29:

ABiological an# Biome#ical A% ect% o$ Genetically Mo#i$ie#

5oo#)BBiome#icine L @ha!macothe!a y) 27)-; 20-/1;)

7. Chan# @ K'Ochatt S J' Rech E L' @owe! J B' 3a"ey M R 6-7889) Elect!o o!ation %tim(late

lant !egene!ation $!om !oto la%t% o$ the woo#y me#icinal % ecie%' %ola!i(m #(l&ama!a L), E< Bot' 07 -.E:/-.:1 htt :HHwww)molec(la!/ lant/biotechnology)in$oHgene/t!an%$e!/ metho#%/in/ lant%Helect!o o!ation $o!/gene/t!an%$e!)htm
8. Chan# @ K'Ochatt S J' Rech E L' @owe! J B' 3a"ey M R 6-7889) Elect!o o!ation %tim(late

lant !egene!ation $!om !oto la%t% o$ the woo#y me#icinal % ecie%' %ola!i(m #(l&ama!a L), E< Bot' 07 -.E:/-.:1 htt :HHwww)molec(la!/ lant/biotechnology)in$oHgene/t!an%$e!/ metho#%/in/ lant%Helect!o o!ation $o!/gene/t!an%$e!)htm 7) Ch!i%to( M A) 3!a&e @ M) 6.;;09) PGenetic mo#i$ication: The !o#(ction o$ !ecombinant ha!mace(tical !otein% in lant%P) Nat(!e Re"iew% Genetic%) htt :HHwww)nat(!e)comHn!gH,o(!nalH"1Hn-;Hab%Hn!g--::)html) Ret!ie"e# .;;8/;7/-0

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-;) 3omingo' Jo%e> L) 6.;;:9: ATo<icity St(#ie% o$ Genetically Mo#i$ie# @lant%: A Re"iew o$ the @(bli%he# Lite!at(!e)BC!itical Re"iew% in 5oo# Science an# N(t!ition) : :.-/:00)

--) 3omingo' Jo%e> L) 6.;;:9: ATo<icity St(#ie% o$ Genetically Mo#i$ie# @lant%: A Re"iew o$ the @(bli%he# Lite!at(!e)BC!itical Re"iew% in 5oo# Science an# N(t!ition) -.) 5a!me!% G(i#e to GMO%) Ret!ie"e# 3ecembe! 8' .;;8) -0) 5e%chotte et al) 6.;;29 N(cleic Aci#% Re%ea!ch 3NA/bin#ing % eci$icity o$ !ice ma!ine!/li&e t!an% o%a%e% an# inte!action% with Stowaway MITE%' C) 5e%chotte et al)' N(cleic Aci#% Re%ea!ch 006:9:.-20/.-E2GR-S -1) Genetically Enginee!e# C!o % in the 4S6.;;89 E<tent o$ A#o tion) -2) Cammon# J) 6e#9' Mcga!"ey @) 6e#9' ?(%ibo") V 6e#9' 6-7779) @lant Biotechnology: new !o#(ct% an# a lication%G CTIM) .1;) -E) Ce!be!% K)' Sonnewal# 4) 6-7779) @!o#(ction o$ newHmo#i$ie# !otein% in t!an%genic lant%G C(!!ent O inion in Biotechnology) -;: -E0/-E8) -:) Ci%to!y o$ @lant B!ee#ing) Ret!ie"e# 3ecembe! 8' .;;8 -8) htt :HHwww)e%cience)w%Hb2:.HL.;HL.;)htm -7) htt :HHwww)molec(la!/ lant/biotechnology)in$oHgene/t!an%$e!/metho#%/in/ lant%Hli o%ome/ me#iate#/gene/t!an%$e!)htm .;) htt :HHwww)molec(la!/ lant/biotechnology)in$oHgene/t!an%$e!/metho#%/in lant%Hmic!o !o,ectile%/OR/bioli%tic%/OR/ a!ticle/g(n/$o!/gene/t!an%$e!)htm .-) Key In#icato!% o$ the Labo(! Ma!&et @!og!amme 6.;;79) Inte!national Labo(! O!ganiFation) htt :HHwww)ilo)o!gH (blicHengli%hHem loymentH%t!atH&ilmHin#e<)htm) ..) Kimb!ell' A)6.;;.9 5altal Ca!"e%t: The T!age#y o$ In#(%t!ial Ag!ic(lt(!e , I%lan# @!e%%' =a%hington) .0) Mo$$at A)S) 6-7729) E< lo!ing t!an%genic lant% a% a new "accine %o(!ceG Science) .E8:E28/ E;) .1) Molec(la! $a!ming 6.;;89) a!l)gca)ca / Lib!a!y o$ @a!liament) htt :HHwww) a!l)gc)caHin$o!mationHlib!a!yH@RB (b%H !b;2;7/e)htm) : :.-/:00)

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.2) OFt(!&' et al)'.;;8 PGly ho%ate inhibition o$ $e!!ic !e#(cta%e acti"ity in i!on #e$icient %(n$lowe! !oot%P' New Phtologist' -:::877/7;E) .E) @LoS Biology / 6.;;E9 J(m ing Gene% C!o%% @lant S ecie% Bo(n#a!ie%) @LoS Biol 16-9: e02 #oi:-;)-0:-H,o(!nal) bio);;1;;02 @(bli%he#: 3ecembe! .;' .;;2 Co y!ight: Y .;;2 @(blic Lib!a!y o$ Science) .:) Re o!t Rai%e% Ala!m o"e! *S( e!/wee#%*) Ret!ie"e# 3ecembe! 7' .;;8) .8) Sta#le! an# S !ag(e 6-70E9) PGenetic E$$ect% o$ 4lt!a/Violet Ra#iation in MaiFe) I) 4n$ilte!e# Ra#iationP 6@359) @!ocee#ing% o$ the National Aca#emy o$ Science% o$ the 4nite# State% o$ Ame!ica 64S 3e a!tment o$ Ag!ic(lt(!e an# Mi%%o(!i Ag!ic(lt(!al E< e!iment Station9 && 6-;9: 2:.D2:8) .7) Sta#le! an# S !ag(e 6-70E9) PGenetic E$$ect% o$ 4lt!a/Violet Ra#iation in MaiFe) I) 4n$ilte!e# Ra#iationP 6@359) @!ocee#ing% o$ the National Aca#emy o$ Science% o$ the 4nite# State% o$ Ame!ica 64S 3e a!tment o$ Ag!ic(lt(!e an# Mi%%o(!i Ag!ic(lt(!al E< e!iment Station9 && 6-;9: 2:.D2:8) 0;) Tac&et C)O)' Ma%on C)S)' Lo%on%&y G)' et) al) 6-7789) Imm(nogenici#y in h(man% o$ a !ecombinant bacte!ial antigen #eli"e!e# in a t!an%genic otatoG Nat(!e Me#icine) 1: E;:/E-;) 0-) Twyman an# Richa!# 6.;;09) PMolec(la! $a!ming in lant%: ho%t %y%tem% an# e< !e%%ion technologyP) Science 3i!ect) 0.) =a%hington 3C 6.;;79 Biotechnology In#(%t!y O!ganiFation) htt :HHwww)bio)o!gHhealthca!eH m H$act%heet2)a% ) 00) =!igley C)' Co!&e C' an# =al&e! C) 6.;;19 Encyclo e#ia o$ G!ain Science' "ol) 0) ) 0.0

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