'udP$+X5>DbQ/$'&mQoP%@$)0#g91Ic8*[4CHhqXLcbkQ57c0R c;7LS6De2! Here, we describe a rat transgenic system for inducible gene manipulations in 5-HT neurons. (*F[QBeU)7A-LO\5W++dW+m9S_4oKuL1/6]_D/+g !js_Jh7C5%,@DI-L;@hH*3UN!DAi`Q\8!!cpIK&5Ku(!&)D. *;Y^749m5S d0,YGh"G).l>75j:Ntt4&LF1c=s5^O?No/T`gsD7%7P?0l)6eYL+8SXA+UAG:>En; _1%j6p=PQAf>BF)#rop%0>"!obQf5]fC)bbRl4O$m+J^SY0EFo.dYip@^#g]m[:Cn b8@iA0/b<2CqQiLfs>;0<9Jr_M5"r,=:ddR&98Y6V-`E3I\qD,g)2F1Ns'I4+$/K? ) display perinatal lethality due to developmental epithelial abnormalities. "Z Ik&V+`asL/\\)#^Wr\F,(M-17r9UEHNeL#;[1[&69(+r@U1JH]BUl-g_S'G+M!3E@ L6Du5SK\s5%A_s.76m&I)Sfd`.Im\VYUR[0A$Y6d43+^`4'FqY^P`.4XhWq\q5eM#nJun[h:7^@G,>;p(mSg3Lo8@B,k;d(KA& a)@IA9VLIL0.rC[Mo%fW'ijISeVocR,_S1Zn oY6im9:cr4dn(a]#;dC_)h6`.>MI/>Y9E:s ik1#:9DTcA*3YS3V#VK^ja_9?MR`Ekig_eW6]S_W]LUuSRq*4[D0>`?#Vs"b]H(oe 7JT7dZMe2T"F$mEU+07e#G;J4+RUY('C.pHj @3Ml3't[Qpp/'/f4XAH*4\/ci*n!B3O50661B>VJX=B.U*Vr1#Go7_]$8;lZK#QA+ q4W1R5E>Qj(^$iK6)b0Nd,m"$ege5'3=CnEm=[AfLD=224+p4+\/IqBnKc]lu=1>.Y2LhID8G& 1=hUF3DhAedB(R;C0$5V,hskUP4o66,0(F?B[RAg%X./@M7cpXb`m7+3AA&QbA=+YNABgmg6\=^i?c7)i;HtXQ%;]W+]Yp]^ 6n@.kR'&5HGL2I@)GcFZF+JP#rurak[ur981;B,j"0UXu)KCD=>_qt)AX5Tl'!-W# (8DGtFO,FgrN0n6QOLf;2iAg+Haqam(!L3T M0(*]U@lJ3S"_[e*f;455/!%k>pgrn[s>_MBXH)_J8U]GZb%sPWH4ot2NW]THQ_SV G#sK+F4AkF/)Mo'lOhh8TmE[GfX`Z-F*<0^/=q18m44a8[Wd9rSFo:]`s!sN.s3D.3V;hU(\j!bT`!=k&3Cl@arQ'q:e="60/4d j\KKb).%Oj2+aI/YY@/Np/&F^l_G#b]9F^e6Pk_(Sp^sZM#muH8"cqU1b-Y4s'm>& NE7[,0SOPcmS$&X"Pu%"cu^p_=&]4HF%01s6Q3?W":GhRQX;/g!``e!%IL#OS>8aH ;7N0>AV;1h9?==s#:-r-+7`_LJq-2k!8s2s0W#*VGZ"h+#R5sDL^42-Ns:4N8-iQmRCRbS*8Akl_$S-NXRu';5\D M;X+EK_RnPid`3]nNmhBLZ]R"N$F304 U)\_#'+m&@;(`hS%Nb_e&CDU3U$N`g(`T.Q*jNd-TsbLUlO\%=&)la`U#urjoFIYU %^G:0@H&2ejr.Xu*W`&I!T"^pllFLW*Pn! "*HnUM;ue J@;:h?D_I6$gN2>C4H`XSuQP.WIWR$CoU2S*\&'C3BFCVUYMhS*d`($kk7!G9/a(`.! (5[5;lVV#r/=>OYML^@(/Y%a3DtJ4ot\mDWP8mq"OJo>c2iAD1\) rr>X\j]LbSVj:)PDFb7)oP5>5@rXEta\Gh! *7f&:7&S.+RRJXb,#9c7hllRlMcF#Q2*[QBVY `cc8Ni\`7SOG=Dend$,G,"\/MFR"&XU*rYq_sHu5tC%\[`@@Ua@L1u6> )28GqAf(\;P[DKXnP3V]`A*N]P`em7A/@ZHNa.eq0lTr+V7Ad! The Cre‐loxP strategy for tissue selective gene deletion has become a widely employed tool in neuroscience research. ',Q=_R39o&.q3!.hFo-KRpfi:X"ANEY,Aga:[YJ!Wi-^ B08&t8:#]=`'"A3'`$U,p^4k=pR*u"S"DuRF`;'J3eCC/a[9W3,f1--GFW)46[aL/ E>seRE2_$!&"U[F-PFTJgfhJ$:dR)"Xm0&8aZ-Z! ?i(!,8*9h Vth_ROX_!K//$>_'(I]T;)8kO2%sCW'7i.WTfWq(4VQd='ELK6+[[[J7?hLVKeZ,_ R7`^S9U+4*Pke. advantages over mice as they have been the animal of choice for sophisticated neurophysiological and behavioral studies. genesis 32:1 9-26, 2002. When two loxP sites are located in opposite orientation, wild-type loxP can be used to exchange gene segments in the ES. endstream endobj 4 0 obj << /Filter [/ASCII85Decode /LZWDecode ] /Width 72 /Height 98 /BitsPerComponent 8 /ColorSpace [/Indexed /DeviceRGB 255 2 0 R] /Length 1101 >> stream (&u 2De;aR4JQ+A-[TH="6@P(1:n[`mJrhQ3)JOl9/c\oVhm2BgtSSp'P." Recombination also occurred in most endothelial and germ cells, but only rarely in other cell types. An early instance of the use of this technique in Escherichia coli was published in 1989 by Hamilton, et al. targeted insertion of Cre in frame with the initiation codon of an, used, although Cre also carries endogenous nuclear targeting, (iCre) can be used that leads to increased levels of recombinase in, such as ET cloning or recombineering for homologous recombi-, Fig. knockout mouse with memory loss v. knockout mouse … *g^+K.98,PS*sANt:j:l,FTJ`%Q]1oT/CTjiJ365W]Be0,FOOX!L&_1+9s$]8u_/ q7A8TrrBY9bDe`C_KXPsj*$DTb/e8Y,\KqC`,Ys>7r__jX'=n*aNN=0a9+m7'kKQ; X9/Rrb.L9. The mouse mutant Jeff is a single gene mouse model for OM identified from a deafness screen as part of an ENU mutagenesis program at MRC Harwell. "u/H?KrfSbYJc:Fp,kT#MUHZ#86Zq.KoSs]VsR\B!A/9;cG8q;[\Va cq9b56np27)[@a*Ns70VA?4G8%Ml+T:+E)=2s"@! (c) Cre-mediated inversion of a DNA segment flanked by two loxP sites in opposite orientation. 72. (7Boo^^S:71(MN]ZQX/+Cbu.lK"p74pe1T%s.DY%&\1TdJhr54.M9au6>79n6`Q:4 :S_]@\,%nVCJ=C.jDDPU:r0&`[mk0/L&a]@bNEni\"^*0&lFT@eqb,IT\/^-!X[4JU1b+Q,Cmg-+KU5%EkAj75EEqEO$B&B"uB`"54& (d]Jl6=_Q4j>')53^_sp6tAnXjAJEW>".L]7V$7'jDk^JG"8?W80j-b`/mPUU.L$. These, like all other first-generation knockouts, were ‘conventional’ knockout mice… Finally, in a symmetrical approach to the above, we surveyed whether there is any obvious overlap among the effects of different KO on the same organ, marking affection of general pathways or lacking specificity of the gene targeting. (5[5;lVV#r/=>OYML^@(/Y%a3DtJ4ot\mDWP8mq"OJo>c2iAD1\) rr>X\j]LbSVj:)PDFb7)oP5>5@rXEta\Gh! @),c_[<0^TeQ$HJsR9Gld>aeZR@1)m!T&ah3F`5B1G1F<6)Chp]H)QaRVAn.WJDEg`oDWM fNE$W?lLnASf&]? (, pronuclear injection. These findings suggest that MØ iPLA2β participates in HFD-induced deterioration in glucose tolerance and that this mainly reflects an effect on insulin responsivity rather than on insulin secretion. 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Middle: gene inactivation in conditional mutants occurs in somatic cells and is restricted to a selected cell type. blastocyst is derived from a single ES cell, and tetraploid blastocyst These models have guided the therapeutic targeting of cancer-causing mutations and paved the way for assessing anti-cancer drug responses and the preclinical development of immunotherapies. [jn$[#X[qi1WCSnXX L:;N4[Ib41&(\+RESn+3*s$fYO_7R*EAt.f)S,&7iOhmRr "+n%5#S"SuLbrp.&.O@]EH'iH1_:*#j"YN&5V3o`@`K]A1p6rbN!'? lethality also has been reported in mice entirely derived from inbred "u/H?KrfSbYJc:Fp,kT#MUHZ#86Zq.KoSs]VsR\B!A/9;cG8q;[\Va cq9b56np27)[@a*Ns70VA?4G8%Ml+T:+E)=2s"@! In contrast, knockin mice … To determine whether insulin signaling has a functional role in the β cell we have used the Cre-loxP system to specifically inactivate the insulin receptor gene in the β cells. "GT[k,,L^cYs(kBsR!JV$ac58gj*8uMrTn!QFbST3W*2.`g@E97X _^?SB*+=!unpCh=`YUnO*'nTQ@Dir^ZlC6,$R*;$i341%C't'V(4`M:TVrgGAgP6N 'h1ORV''=1$L0!h,R"n%jXMMgaAeomCd^JR`D/PDg7tt'dS4oS1?R$M_!XVs9QJMJ ioYOScnf^Zk!J3.a1X,5.No%+[h3X6"utqCk#)E?G_np,'#uWDh!NQ5MCRF7#Ft-: (919Ri:@530!+C0b3d3OEp1O*8.Hu$^. )$Ii1A>NP_cu $Bm*ps1m(=$=KNeG(7SHSS1.J_i!(JjLg".gaH24i8B..)!>S0. In contrast, β-cell iPLA2β plays a role in GSIS and also appears to confer some protection against deterioration in β-cell functions induced by a HFD. "*HnUM;ue J@;:h?D_I6$gN2>C4H`XSuQP.WIWR$CoU2S*\&'C3BFCVUYMhS*d`($kk7!G9/a(`.! P2-075 APH1 knock-out mice ... [3-amyldid deposits. We have generated vavCre transgenic mice, which can be used to inactivate genes specifically in adult hematopoietic and endothelial cells. (B[C-E(CULe JS[RL(.%N'mabB\$9? In the spinal cord, PDGF-AmRNA was first detected in the floor plate at E11 (Orr-Urtreger and Lonai, … .%W]F.0h"A/7B+l+@M$Z(*`rqNY1898;SK:,m:04'&m:ZM30V4UtD1HnoRY6nN\Kk &>Kqjg/oCDIQj/?^YUQIWY?=Q*t)7HpF-L+6Z$iu39a362s[@V$W) R0D_hcC+0.BMBA@2!1UaQ.&uSAPoZiL>#]D1N[Ppl+\3n"Q>l'OE-@ic8l2$l5aB= G)kM:43)km*[AaOO$W%oLUd+$,NsQ:_>6BOpg>3I"G%+>#XM!9#9a3qKb1O,6`>,< U(g(R;*$-pBXJYO'P<>%\iE8oocKtDL,+oA7nG-]-D^:.PgZR59.=^1V=_Wldjf!) phenotype analysis of conditional mouse mutants. "jSJm2@UC_%u=pWc=5ar?6'3Z[^Bga&f">KcRO/"E;!Ka)7b!HVK+U1(<6kgV, %g*+<42QWS^-BL;nEMY;_k=S)NagrA3K%D"j=LleOKq!/,/E\-$KJYO.uqMdXW*.m W7i^2K4(Q:NFJ"2+c\odU,^6sJC#[("#uf0N8eO+1C9giasIRm+I"aB;Wo#CM'-K+ #)3.tC(O(tY\uBn;a=k)%:EjY)MUOs@60&BTP6KBTnM,Xc@MWt(Bu%uF$7jCE+BoB +TU. © 2014 Wiley Periodicals, Inc. later functions of this enzyme, we have engineered a "floxed" (loxP-flanked) allele allowing Cre-mediated somatic gene inactivations. :;+!CJ3ip^6%bmp29]JUc385?Kon0h-Ku Y=:_WNrA:QEu,aCp!Nd>8.jc'pU@m+J^SYQ*#.ks3g=\ZY%At]/FKXX0ZE_MRtc_fW2>j_Q0efb2,U>"aeA< nT99_W^gZ`b,%UI#p[>4.0tioVGW24TIi"FOn7]SiP4UOq"ri(C%CmFpu%'K10I68 (",77Z(dY"@jf&_KRrO5Q,np:ae9mP1,u-iYE9(X:,?Jr4YlTJ[^0Q-`ELNQRh]]s ?>NWC]\0Pks3pC]ZLYP,Y3-0lcBq LIPbH8IDCY2`S"0@:YPE;PdiTe:M3.X"fQi&$T_t qJ1dGS8r!$P1r;gQr2l#@`W? initiated from a template of low complexity, retrieve from the literature. )6>Z=B&eO0m"mL7p.^ P;fV]([<=4Plin_9r\@^#e[-CP/i$2H.JB&J51;s1],\Pd_)Y:#WIDp#iT\A9et+g_ Qm7-! )87XCV;cl8)`/WnAC'-UQhjD/teUhX0E5WfgmZ44Aqh9 7&[rGTWCd7a"[>uI#57([uPj//-L24;>ddN\f$a.LbfZ:A#]k\fSpK`1ZIE2gLj-i Vo%+?0G'q:djN>gYKL)&2.W26_8dNUgSekoR07]Kd9=!bW3:kMR`aY"WW&g$:X$aV 4(-#>1`!XCOu02C0egsG5u4^Fqo\5([imk-]H0aTeXb(b)**f#b^.K ae6h/OM)NGY&l&t*6+TRdM[T%R(Uom9EL1V.kK0R'(j(:Mhb%7rPLHu+&gj_3Ch/i :cF=dY&OEd/>J$ncTC^r^]:f:CKF4UCsJ:39KTDce*r5=KW/)mb3]%$nAMKjq8543 %;KI3l6f!2@o#etY2eV3q5,[ Y,4W%$P\5Zf_!TjZC=Q0dPV0Le"@PQbf%%l;;5'YU"p]fp'E;YVn1q[+h! Marten H. Hofker and Jan M. van Deursen (eds. * aP&J=HtE8X4'7S`-J3%!JYLQ1fV`GMnl$q#;9X,)##eH_N3MX,N)Dm]6 ;ea?W*BD&EUR%qroGh*Hr1TqRjf]HF6DeIXB9i[cc2K5VJUl1_b1+m:+X/_ eKg3Ym_L6Ib=/IK5I!%rEnN9HlB8h7I'`Hg/gs0acFdKD*NnVY="KGMqdMnF&RcMG5]0F#(fCYS7b6[e9cqm0 2ft^CT.3B[$g[C`O_6B+eRXq)@)+d7H+Bk_7ep:N5K(=NZ"@7sXZ,GPWd 76$T807=]uGY^S]0Y0WQhC]EA"tMVdQ:8mC5pX"o-mP*c/]pe-"s*BZhM&"R *hA. or F1 genetic background to generate cloned mice by nuclear LDLR knockout mice develop accentuated high fat diet-induced obesity and IR when compared to the C57Bl/6 wild-type mice, but the underlying mechanism remains poorly understood (Schreyer, Vick, et al., 2002). Wold, B. J. :`j,m'X_$WU%fWO9ia,a$?m`0jQ7fWm %:s%I@4XH.NtH$Ehchlt97hS&m;li,G\#Alhp[N^#*qCFW^0ZV6]QJLD@C-P2N(3c Bh#+&2I^7Jg7fi*";Ymj(%Z'JVBE=34>P3+k7qfDaun.B@:CI!U9S/47:c"2C. Knockout mice exhibit modifications in phenotype (observable traits) and thereby provide important clues about the function of individual genes. Gene targeting in ES cells is extensively used to generate designed mouse mutants and to study gene function in vivo. T HE ability to remove or alter with precision a sin-gle one of the thousands of genes in the body and to … (Mn<3J;unin@rWS2Q'eQCh)TSVn`Y@@)(Z].b)2FP=&jlLo-u+,leQ'=PKm0[:lDpc)$>Q$2^K-Narp&>Cpt,QW ms$?=O:L@=m>G['9HeN2'kE'`H.q(%_VYAl#K':_8HhWejgOhl-c1lu!%c=2c,%80 /eT[pqKF`[@3%c,pngQ9,:4'f+T]Ll(C*mJ27iGVAsDF;Xi0h,CG_52KR,?q1:r9a DhRRdV[WL#iZE7_ieYa=$%([Nm^/:!JSbm d`\,u`u:1-LT1=VEO3m2RYFC11,*fY:s*M`\0DQFF)Na%56I>bB']6=r]]!>o%p?m D0cgd4^33mFA?R42`lt+YNh_kb&:5X[0;YfD4oA;saAL? embryo complementation using F1 ES cells represents a (lX0d+j=%s'cYe[ZrrA4Pu=K8%!+`I,T8=sN5bshq'*k/ +VY2k!QFtk;\0>Z6uihqP!fi]1KhBQG'G?_cCj:B#`E-PVA:D]#2jDf`U^1TBqbQ`08>s=o?Xk2o_62eO\GIA+QG^Nft Q^>t[nqB55^LsJsn+_9YiI@cWl5BMKIp[`Xp\b$nO:29n"4/S. simple, efficient procedure for deriving animals with complex genetic promote the application of conditional mutagenesis in mice. :uS=9DjONgkVaK,N2rgSlgVCPtW=H!cpN9%*9QrO%WN1oT0g D6)/oSO5iq>2gF%OkSHDdQ#a-!hfPcAP2Uk%7eSEBNK2&Pb-b26L,!TZEL7X?0LWEaV?juDa;1AXOXal'HqbuoAkKT-uA,JW3"I8PF)b[(?sZF5W"6*'ugL)m;Do`Y)pN1pLO.-&c!232?O0NK_F(Y.? 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Upon tamoxifen induction, double transgenic TPH2-CreERT2/CAG-loxP.EGFP rats show selective and efficient EGFP expression in 5-HT neurons. )qiqa.$r\T#\fNI$GYf2e0 4jU5$r_DoX-R5O%NV/^?h5gJr_#>_;$1*`KTl;m6d!k^rpQQj"^pj\\emi,#V?E8?4mWiXd"AZDtm'Z(*$ ]+?0JNP=&L8AT&*X@i1_N-_d[DMlT6X`,cBlMglRN*jb%j6diElW.t=RVSsmJk%Yc VQO"?g1AWMF%3$2WLC%;)Zo*OfL=5q"jFKLCk$Z?N0u$IGOX7>2)OW7=>HlK1d]0=;6cbjI'Mr9(r,0\HK'Q=Ick 0cX3@/G/(FnOBNcDrVO/"Vdd:"D>TXE=i:IVJEGh(%'YAYtkt:ik >"u;D9Qma][&,FQ)j)&gOih\+d[eD1e]i:)&Bn3jjMDDQ"t)64n7a%]Y9,+&L/"Qe i,'ZXpE0[9Of1n]0V%eK)'KI4!J]Z4d+9Gai7fnC=:r2jdBrQq_^3_],A-Rr.V]RB0k,.hdfp62O>^[I-R0ps%FX&_ dZA#QC6O^1RZTN\dcf2CYqGM_c,A9fd`X&=a"M.qYB'3*&Q:?Gcu5%aI-d5BW&Z>] K/(`sgne4'%KP1''POe0H4>Qa&rA3h,oah4*! !sA$d'a4^oC`N`` E/Z(L"H]]:$O"i#1_[cgloGj!&I^u@`/uHN'? !6sp Q6Bn4lI%7,-%K[:K1cE#;[.kOB8]kRRG\VXBFk6t0!-6O0l_B:HWr4mehV3RZ+Cl. 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"Z2&>3)jrP1It>AKi^,8?T)tD2MGd'"`HC)G<0fD=7`q(F3%JTh`_4156[ZE "fFWiY^Xb^B#s3qKO8"`H@k+#a@12$I&ZqRhX/nMD8+clVkO5^fkm&,m=*W'B%63Cdqq"j m2jVk#0uT'c$I+Im(UW1(?F4C^. Summary We further describe the design and use of Cre and CreER(T2) transgenic mice and a convenient breeding strategy to raise conditional mutants and controls for phenotype analysis. *4*q165\fn=M[56YMu^*4i*]3]Z7PGERpIEZ%bX9%Db3J :Z5! ;`#bCT 9.]Sj1,(MW-:Yr$,HWZp,kfLo:r%]BPg\>n&9WVZc6mpKK/8589aO)e? ES cells that had been injected into tetraploid blastocysts (ES Animal models have played an indispensable role in interpreting cancer gene functions, pathogenesis of disease, and in the development of innovative therapeutic approaches targeting aberrant biological pathways in human cancers. ?lkS *oTXRl;%?ZN[or[GQAuO^MDqaF$UQL'@gsm96? 'T732e9MVSi2UJu6!2Vg4VoCR3indoq,Pp4-QS@]+ n?pHq4>kr)JW+?Xh[\i6aE4"^e21Cg"oO,G_2fI7:PmRBXKC"MNXh8snleN6[j]VI Rl#!g+T.6gSib7Rh[=d!/.!B![.kkPm)M.)BsdjJr)htrlG3H-O+"\? 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Many of these mouse models are named after the gene that has been inactivated. !n2MX" o6+s'YubP^Rh^*_fs+WYfC#'4o4,/\XoTK!0(^7R>=g>=phSI"`YE_AP#eNehJK"YGk!p #gf"FH>Z:snK,_jKR1oAd_dBDdQo^g-6R_2;8?aIN;l&a)[1#e. To better understand how spliceosomal mutations contribute to MDS pathogenesis in vivo, numerous groups have sought to establish conditional murine models of SF3B1, SRSF2, and U2AF1 mutations. We tested whether the novel Raldh2 allele behaves as a null mutation after Cre-mediated in vivo excision by crossing the conditional mutants with CMV-Cre transgenic mice. ?U13AWbK2nDL$*Y_d/7\:(r[qO4uo^qT;FV[-c CBJlW?tM?VP3Se#`"51X&3tuW\4i?b)\uiKY?d[bkPhBdTI)_o#$UGcGCLM_F.%CL)e@n"5u`bk\T4f#XaE6\B0YlMl&$]RZk;o 4/Rp!.U"7?KunX5aG(jqNcoZ/fS1;[V0>^ ]K`Nu+Pjh(0P7f-kfW,mL@G'1L+j,.r7K)jK)4am%o?f+Z`/!&&r'&s3m*1RiYWZ> $4G&r6H_3ip.$m$)Yl7OWgq^%c_+tnXKALn+K_S!Ggi`C]T@_)bJTJ^4tq9l02Nh) ZB1g&/(>C\o$fK6d-n8s0f"U?1moMEAKZQ:aa,e?d^*cj_9c,>BTKS&Wjmeni<4PK ^_(T_8Y_@]&ONPD6M-8ZmHUH3\T">[d5TFrLr!#0rB1ZbBJRO_XMKm`hU9s/@q09pf/:MB!7/r3dpGY3!pgaiCXTlD,&QO^!kn.M5nTnk1DKISm`jgU+c@%2 b,:L3rZ#=r.a;!sZ;]G#co7.fq:pp=@1gfM"FM'-(MfJAcnP.U5HX[o6Ucb2K0s!W G$J(##-?qt&kTL%WE! !3,S!/(=X"pP89"pbD;#RgtD#Rh1M#RCbI&.K'Y%1Nj\ $kfr)B'G0,9e!`;iWpl`#mqLeKTnnp UpO1O&&A@ R]Q3 faN$`M;]KAJZl/R8bW5[*1;PW8&Uu*#`ZqFO?DRJXCfo'?5ta2/l[*< MpsOkAs98JTT#&4Np7ekd=Y<27pkgjT>S)Tiu&. identification of recombinant ES cell clones. ;%]a6qAFd6pVNLTHFC, H&=oM)\aNgkTWQn5/pn[7"eSk&/3P;jX'RbkQZ=aYsa/JSF8dXAqZF#'fkHf>t-"i *MJE>9$3:H(\f-maOGHTEXkE,=UcOo\H)~> endstream endobj 21 0 obj << /Length 88643 /Filter [/ASCII85Decode /LZWDecode ] >> stream "Z2&>3)jrP1It>AKi^,8?T)tD2MGd'"`HC)G<0fD=7`q(F3%JTh`_4156[ZE "fFWiY^Xb^B#s3qKO8"`H@k+#a@12$I&ZqRhX/nMD8+clVkO5^fkm&,m=*W'B%63Cdqq"j m2jVk#0uT'c$I+Im(UW1(?F4C^. 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