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These results suggest that Cd6 is valuable as an alternative site for targeted transgenesis. Tell us the name of gene you wish to knockout and we will design a nuclease-mediated strategy for you. :tqUE-=[)=cg:) 9O`^@$CZ!9)g'tRf2i,n/`i-1Ndb5DJ5MD[3GmC,A>=&&g7:03pkq)CdU_!dViZbqVlc5`na_F/:B])UP29pici/]6cdIIVWt?iJ^jVq[9``t4mp*ReZ=D/= _i>UQ__E9R?hr:5cOLB93&:!L&G:n"6k4^okOV"8G=GemAY2nPA.Cn$Ru]SM,_lN4`2qd6n'k4r8:N7k$8PD1RNr%] "QA3*2ENg5fMLqfZXU-Y< P\lYW@uLOV,]$6B1'o!D/\LE+PhPBu@`cXP1HBD7]W)p.^Y+E#.E-)iqX=s6Rln/u BX)m7BV2k=C1H8J;\,fe;i-QuGZL;N8L4.FmsTFGDM0HJ[&6ZLK21(#(-jgX3"5bH EUq8l$t_bU+DU%WSGEiLT'a:[aomBVA3!+)Ih,@G]4N? s4IA>!"M;*Ddm8XA:OX[! oFV9)&r2QY"p>W+:h,4cGm6NB6$sN(87@:R!J(ZO*eo2nAA^D8Tn(n42U2XtAjVSj`<7Omo:$k,S_@0QiAL&;10oEE5W?j60*"mc:[:gXC\s$gs6 `c,5fe,9HsGD4[S5HJ[TaBQEl15? must include restriction sites that are compatible to sites within the generic targeting plasmid pEasyflox-DTA. [W,)oO:h6F ;L$b4=es>0=:48`+@\mXBj#?7I.>JF(!tJ86/"LK/kl(b&[:%\H)[:,cMC%0XE/3^ -95`)@WXUYSF9aFfjq1XMH10) $)6+A:pYH!dr-FC\e_nVSBBe3,9cR8Ya5h!KE=. )6>Z=B&eO0m"mL7p.^ P;fV]([<=4Plin_9r\@^#e[-CP/i$2H.JB&J51;s1],\Pd_)Y:#WIDp#iT\A9et+g_ Qm7-! ;;DB&hrOCMp/,;R5l /8HiqB\R.;`#TR\iK$Y?NpdXbZXZ\Gl[kFeFiHBWJ5[t1aTTk1ea;6'm,n2#^0*[fe(P>Or6rB=)7_$\Zj'64. ZH+@&o7=_d1)A&ub7tJZ>AP"4j]=BpUYi.U3(@OLD8Tg4Q9>.f:RAZ8-?1Mq__%"* ij/=VCpRt%`8UA,^%Vb%`n]Ql(]%_;))/8XD*8bIUf,*tFkGd=?,%55utd+:@>RSKfUCkNS@/.DrN\\nmi;WG]r_t);H nB1n)6u(c-ZTaCs!4fg_ZZ,ZWc22gW2@n8G]fL)lRh*:goH`?bOmKIa8$_JfpHJr= SEiKs. 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Here we have crossed this mouse with a floxed androgen receptor (AR) transgenic to generate double transgenic neuronal ARKO mice (ARflox::NesCre) to study germline deletion in male and female transgenic breeders. !=XN>03 7t/k-%"oh[6jVejnJoRrW$c>4Z0gEjH`#YGP$RH/s4r$!J(::J!%=!c3/u@K@_]%igp@gg0/fO?Ma%d$@;*ECPm8[+m=4/(8K9 :/ p$4],^9MU%=ZpF1gba(*`ULpmeFM^WBj,:WC2qK4F*_i8XOGrCaR5^Sm'KX5\?/Q8 m[Jm^XHAU+huHe+_Ym_@f"T\bG_/3;. )!/!-K)(sR"Lij2VWnUF?PZY8G,^Ee$_.HmR[EhUX-C]SM2-]2qDVr/I(#KZBDG]))QoKs]4ZGZ W*6O+LG"tuOe'QlWC6K?(A'6ts*H_RFJiB/esW/++cf))I(? This work established the feasibility of removing or replacing a functional gene in bacteria. (1996) Know your neighbors: an internal ribosome entry site for bicis, gene knockouts in the small intestinal and, specific recombination mediated by an adeno-, mediates conditional gene modification in the, Optimization of spatiotemporal gene inactiva-, tion in mouse heart by oral application of, (1999) Tissue-specific knockout of the insulin. @V-sAGRZ1V.$*I2gLX).`>XnQN)=9q58?S:0V#ZC[8]]-<0#75b=[X=#2b>``[+0I6.VU9d!V@$a*4$,[0:d :W;g#V4,3-2dHmp.CJ^)[u0*N )PNd>JXRNDX).m8D./cHiKP8>U)>lK)$-fV_2iG!NGUt=)M0:S@@.m?VAWfq)Bk=^ 2[U[:$6\=@+^u?5iZ]RU.O*k-,@W\Yi^+o"8gNCo-":%(iaO6DC*qq\-XqBLidrRf MC@JI. !riFk2`^WaqZK$Ne'nP0UO89:O4ifpZHpSV5 glF>MW]4Qp_r0,G.QAhL ^*SE#WQ2H0iXaoh@lNY_0Hkk$AgN8t9M2kn9fjC^ jr768#ll-^o4PZ6df''8U?Ar`o>-W6;uU41A:**L'rSd]9RXu->[ubG]>uTHAR=c< ^B`tlfNROV? 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KNOCKOUT MICE J OSEPH A. M AJZOUB, M.D., AND L OUIS J. M UGLIA, M.D., P H.D. The timing of gene inactivation cannot be externally induced and may occur at early stages. J2Q7]3$]7K#D>EP:q1$o*=mro@So+\H .4CA#,gpCkb$c`gc),QmN5?_eQV1iX8W4R-!hn$he>ZF@SN`rHWADXaJnmBH_e/L$ [. :8'MN7a [DZrXOh"=3hbaZDQAlLg,[*D1(()?K3-)5\.GZGRoa9*aR#BqP%*C`\qkX$cR:5e" 9_tL;9eXtM)goZ2%1f(J-6562_P#nEnAnHMAN'>pd]fH5&?M+59gjZjO0(hc!<1G5 i3]HC],A%U3VJ:e32^iY&gC(EBXpU;:.gZWcg\^tR#b1u_Fts$;uk8jj]nOtJ4gS_ -on6U;0c0Hp:#oF)Eb-$S0q!H/s.! oNMA1Dl@F@_OONu(MTd!E/,Mpeu(?j6=&2IEJ):`=*CUP\nG-:Ed)G>:*F`2(0+Em L\=,j.0FG(39XE'FCk,Z6d9N0b&Ojj+SU4rj;DU3XI[tX1?R(bktI4@EH(WM7tkMj 7"-TQF`Ad7HgGW@(C>Ljja-[ADh&dPP?a+,d"WjkF9N%o0#la+4^gki=BHeU;QuV[c[BN1nhrh_pB,W]'lbROG'EAQ R$EOeoT<0MLR0c:mW?gkA;3"mL=UAJ(3Z"R-p#oJ:q.E`p`>:@RUhVsLNiDIU`G(> ;i-BUMN/3^ih#JGW9>%Q/lsE38H4e d't91Nua48h1G@TAEJT!2fiuYj(R(5T6Co%(q"hUrKPm`Km>>0\OV/C@U)V:(FTcg f0Gk9E7/qVNJjisp4nl5/2%2uk?,? A knockout mouse is a genetically engineered mouse in which one or more genes have been turned off by a gene knockout.. Knockout mice are important animal models for studying the role of genes which have been sequenced but whose functions are not yet known. That method has since been developed for other organisms, par… The former does not identify any causal links between two or more variables; it identifies the presence or absence of correlations. Next, the, strains exhibit (undesired) recombinase expression in the male or. *c'5G7M?E,25E_qLESc!/:N( ! tamoxifen is completely dissolved. In this experiment, two sequential recombinations were used to delete the gene. ;RScW3H[M4I9bXsu0&42X>IepSbXXT0_;?SZ8&K%rdR60 "GpY?=f,a_B-bAi'5K`G='O;Fe)\+TL]jm=bFnud\H;XVm)K3hed+KUn8"h]/(;&(I)$F1IM54dA\`ZEJslS$.F%3 oBEuiqgtU@O\o)n_uN#P&3lQ%!OTCS5 !Zo])$cD[(]HX[T>HTsQR?N@=39lADW.Bdq%D\YOEq$9.0"$lGOLja"1*MZ^aW`bM ZmmL_,K,H6O>K$o$=]e5ES?fLWB=)q4>T.5q_0iMofc6TP]M,_bI'76)3l0L!HA1G R1HR'J0O=0!Q3J5fu9*tGeOgH.:Cdh(n. )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*! :R#ppRJ;!De\;:Fg+0n0GMK^5gZEbIk&97$4#`pQf!&k97 a"K!a$G;l:5W/(Fb. .BPM*o\uCiWco3"jZTNkrh*4-O&66-74?+pU_JuV;T`EB.E+EJ'b[[@1Rt#:hfRCjRE0@_8?8N(=iS#_qXX!AZ9imtEdPs8IF-EXGiRc'LT1^7_T(:!TbiRg[T /p[:lT[E,dZ[In17!$H=K-Ig,ZPZt66YI#'e[MTq3BL$F0F3?/BQqL4d /]Cq4ee6hBb<9eF]u%5$>33#s]%?-.0J>c]gcHG!$mh$=(\7.N[Pl1ng>I"[ ENQY2?X'QsF^IS?cad,'bnp%s^"0@tX^_FS(]2]=o$Y0 i':u!JYU9!Fq[9KUD.3[77_2t2^mLJ8VmiBdUcP>HfDW`=rg'rWV)g8Q9!ZnD?^*N3("6_*$L7o%M6gopIO6O_16KZJAO9587,_X gV(LDA/mnN16*L:)+PNb%,;AFLHs-E_m!eo@GE]"0_--;(jZX$O?NlbaT)oj(Q%KheYLB5O@"-DUUgoNku1!1[BV_R=C+?GNk&fl`]MW[;W4-!_b*X=BX %m:=a%Ug7sa#K(iSLcsbE?EZJJ:8L:9QQ%%r].oYJ/n\b5kWM-3Wu+.ls5]FLdBB6 KIH'tiZ#<3ERH0:7rds+A42H?1a7]GRh9//9o.*m,=bS#n3V4e%\Z!0a&&Du6@;?! 'YZgI5]o@G`90)pgZ/g1'7YKgQ4H0\&jtoDSQ7DTPfs->$-_8'Ip9%5bJu>7H0tP .jc'pU@rrE)Lm[:Cnb8@iLCtPepB$H4F.k`e1 eni"OX')b"@]a7cc&/ou2JcCHZY%0^Rl4S&eni"OX8gc=RBpMK(qV\BfW2>jc&0+k ;_UI>N1E1l9Fiebt%Fd>kaQ33l`hjBd8%jB2DCF'7pj\=WlP"bYm,a-nY>#E%RXo].! :`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. The mammalian models of cancer utilize genetically edited or transplanted mice that develop fairly accurate disease histopathology. In an era generating unprecedented quantities of genomic sequencing data from human cancers, complementary methods of deciphering the underlying functional cancer genes and mechanisms are becoming even more important. mice with non-typeable Haemophilus influenzae (NTHi) a proportion of them have inflamed middle ear mucosa and fluid accumulation in the ear suggesting that the Fbxo11 knock-out mice are predisposed to NTHi induced middle ear inflammation. … 8d(q^KWIfLZqH(b9E`:70F\(;AQOY%O7D/1D?Af$P[APoR*.2B`GLEMiB9[R[`$mS 7HGIbA:fgK=CoYsWn(-g/;&h?\aVhB:a4WO5Ru&-VMtjY\B0WYY'k0p@VM30W+8f9 KSc(Q"5lEQ?lIonV-7IDAMrRO"b`NS3C>Sf:g"3ZWsMeq1f)OR=_rL\#\8WcT&=jr NK*@q%>D@\@(; p+p3%4(ItOA9LUW*"-@A@_Kmr(,YDkW#+pB,AY$ZP).+G("F!Ycf0*$PG0 ,(_?j!]2-R4=#,+,f3Q&JVkb*>UBM_-@oJM_JgkhGUL@Y-pi_G3/4.E/_U'l$fIatu/jebS_XK3Fpb0Mb0LH+"_[nOg,Vaok13]TN__e! )87XCV;cl8)`/WnAC'-UQhjD/teUhX0E5WfgmZ44Aqh9 7&[rGTWCd7a"[>uI#57([uPj//-L24;>ddN\f$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%$P\5Zf_!TjZC=Q0dPV0Le"@PQbf%%l;;5'YU"p]fp'E;YVn1q[+h! transfer. Finally, in a symmetrical approach to the above, we surveyed whether there is any obvious overlap among the effects of different KO on the same organ, marking affection of general pathways or lacking specificity of the gene targeting. oNMA1Dl@F@_OONu(MTd!E/,Mpeu(?j6=&2IEJ):`=*CUP\nG-:Ed)G>:*F`2(0+Em L\=,j.0FG(39XE'FCk,Z6d9N0b&Ojj+SU4rj;DU3XI[tX1?R(bktI4@EH(WM7tkMj 7"-TQF`Ad7HgGW@(C>Ljja-[ADh&dPP?a+,d"WjkF9N%o0#la+4^gki=BHeU;QuV[c[BN1nhrh_pB,W]'lbROG'EAQ R$EOeoT<0MLR0c:mW?gkA;3"mL=UAJ(3Z"R-p#oJ:q.E`p`>:@RUhVsLNiDIU`G(> ;i-BUMN/3^ih#JGW9>%Q/lsE38H4e d't91Nua48h1G@TAEJT!2fiuYj(R(5T6Co%(q"hUrKPm`Km>>0\OV/C@U)V:(FTcg f0Gk9E7/qVNJjisp4nl5/2%2uk?,? ((t:7KRT1E>lOUP=S-l^bs-5 ,.`8]4&cdp4.VKO99o?hn!Z? Although mouse … Jeff carries a missense mutation in the Fbxo11 gene. 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'6*OK/R$p.TZY%I9%aCJ$.dYip @]ee;Rh^GQd-b&CB$2%!RkH9,G&5j:=(Pt)@]XLl]Y;"TfDD^,e&0`7f+4d75KmQKfJjR.q(LDU\]m[1TX6JrKfpG@k+qZ(U!E,(67N[$hLoACa? embryo complementation using F1 ES cells represents a *4*q165\fn=M[56YMu^*4i*]3]Z7PGERpIEZ%bX9%Db3J :Z5! the technique or on the characteristics of the ES cell line. !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&. This, search with the term “Cre” at the Jackson, To characterize the recombinase expression profile of new and, of the staining should be monitored every hour, whereas. 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Chronic otitis media (OM) is the most common cause of hearing loss worldwide, yet the underlying genetics and molecular pathology are poorly understood. In this respect, rats offer many, Cre transgenic mice can be used to delete gene sequences flanked by loxP sites in specific somatic tissues. 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. [o ?.n%I[`Db3/WdBZJk-@\*oc4Sm9u[3X0/R'LC$J:V&Y"baJgP))ts8SP9W aTiT]QX1$tC4dh9$PYCj>qh+S.b=HgR6&f&C>G>nQ'Dg1P4XKZ`Z$F!1^G8#/lg*n J?ih_^tg4^GqXG:3$kR5*=N-q%"45DOCZHM.k$;`@O*=W$N428e8`lmoY.b]aI3NM E(Uk6]dVOmh^i\BcY9@97RR\h]QRfn3Ut/!LEDp. All figure content in this area was uploaded by Benedikt Wefers, tion in adult mice and in selected cell types, a refined strategy for conditional gene inactivation has been, (floxed) gene segment through Cre-mediated recombination. (Mn<3J;unin@rWS2Q'eQCh)TSVn`Y@@)(Z].b)2FP=&jlLo-u+,leQ'=PKm0[:lDpc)$>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? !L1.T11^9YH5Sf(tKaBhq_8PA)L1.l:(K>M$2bo(Ui,)!cUGU$rN\dL27SP%n F%V0qPqbQ/+eWWf#/QjJ*6dB_EZA*0ph)]a28s]E=^dt=7Ta(&EF=PFNC-"a+dcnC L6M[C*/8RS,@B2+,kl=af0opBL(jfq*8=NUQWf,E/YdOti;h[i:D@p.5ZpC-$6%=` SUXE>ZV:a6LDRoZ*7! Like inbred clones, ES cell-tetraploid pups derived Whole mount staining of organs or embryos. into the target gene. ;`#bCT 9.]Sj1,(MW-:Yr$,HWZp,kfLo:r%]BPg\>n&9WVZc6mpKK/8589aO)e? Heterozygous knock-out mice for Slc2a10, in which a selection cassette replaces a sequence ranging from exon 2 to the beginning of exon 5, were purchased from Taconic Biosciences … ;`#bCT 9.]Sj1,(MW-:Yr$,HWZp,kfLo:r%]BPg\>n&9WVZc6mpKK/8589aO)e? [o ?.n%I[`Db3/WdBZJk-@\*oc4Sm9u[3X0/R'LC$J:V&Y"baJgP))ts8SP9W aTiT]QX1$tC4dh9$PYCj>qh+S.b=HgR6&f&C>G>nQ'Dg1P4XKZ`Z$F!1^G8#/lg*n J?ih_^tg4^GqXG:3$kR5*=N-q%"45DOCZHM.k$;`@O*=W$N428e8`lmoY.b]aI3NM E(Uk6]dVOmh^i\BcY9@97RR\h]QRfn3Ut/!LEDp. oFV9)&r2QY"p>W+:h,4cGm6NB6$sN(87@:R!J(ZO*eo2nAA^D8Tn(n42U2XtAjVSj`<7Omo:$k,S_@0QiAL&;10oEE5W?j60*"mc:[:gXC\s$gs6 `c,5fe,9HsGD4[S5HJ[TaBQEl15? 'Xms32'ONU_`6%tL]gQRX01PQVIj[%kXKmjhI@0S6?Kt,34 NT7F"cYcT?aDFH.Zo*>tiO0LD4klBJ6'tofQ>)gug#UUg0ocP`ZINbS,rhgO[5&9BhiRa/C+kn?h.N,3f`ECO5t3+a"-\bK\QiH\I\Li;!9 0tQ^Rf#sQ@g$[0hlocMKIr;Q;/ l8;[Z[I8[C'@^f&1sR&n)HJGoo@l35*)dM7'fU'#DR%De'eauHFkgVdJPX!BdB. )2uobg+".q65e #!E!PMT>HL5kWmQi::I8`PcZc>pERLF=+16rq)EPQ*8DB%dM;gd[iFRe_1CF'nDui 6EA;l77#!7e]`(l\%E7"j:*)IRG5F@nG[f\A@.t3e*iuj(D4h58EA]'BNPQAVVS;D "%;`%M<62pqE5"o@0AY8)TZb!NLNVTR53Ih>M'VSH"eCM-h7!eO0[_H"l8$V%8"msL6l?q7or4DR]*d?%# dmPcF84WMVX`e,Z5. A promoter region characterized for gene expression in, the initiation codon is unchanged. ?lkS *oTXRl;%?ZN[or[GQAuO^MDqaF$UQL'@gsm96? :;+!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? Notably, these ES cell resources can be archived indefinitely and are easily dis-tributed to the scientific community for the purpose of generating knockout mice. [o ?.n%I[`Db3/WdBZJk-@\*oc4Sm9u[3X0/R'LC$J:V&Y"baJgP))ts8SP9W aTiT]QX1$tC4dh9$PYCj>qh+S.b=HgR6&f&C>G>nQ'Dg1P4XKZ`Z$F!1^G8#/lg*n J?ih_^tg4^GqXG:3$kR5*=N-q%"45DOCZHM.k$;`@O*=W$N428e8`lmoY.b]aI3NM E(Uk6]dVOmh^i\BcY9@97RR\h]QRfn3Ut/!LEDp. (*F[QBeU)7A-LO\5W++dW+m9S_4oKuL1/6]_D/+g !js_Jh7C5%,@DI-L;@hH*3UN!DAi`Q\8!!cpIK&5Ku(!&)D. The murine models of spliceosomal mutations described to date recapitulate human MDS or MDS/MPN to varying extents. 'uo<>EJ]KO#Cr&WC7j`ei-;)huba F/ZjOY)Dfm(=Xg'*t'^ b^sr]1Pui:V)?MLSD?,]Z]*rqc@X"%4,[%]oI\C@m/9/,oF1pW:kKJl6]@7#b=0g2 4WH:V'St:2NKgOr2U_bV2,0)fkiGUi"/SCHeX.V 6! Let's say we have a gene of unknown function in the human genome. :8'MN7a [DZrXOh"=3hbaZDQAlLg,[*D1(()?K3-)5\.GZGRoa9*aR#BqP%*C`\qkX$cR:5e" 9_tL;9eXtM)goZ2%1f(J-6562_P#nEnAnHMAN'>pd]fH5&?M+59gjZjO0(hc!<1G5 i3]HC],A%U3VJ:e32^iY&gC(EBXpU;:.gZWcg\^tR#b1u_Fts$;uk8jj]nOtJ4gS_ -on6U;0c0Hp:#oF)Eb-$S0q!H/s.! A large collection of Cre transgenic lines has been generated over time and can be used in a combinatorial manner to achieve gene inactivation in many different cell types. !MRRi[qKljpBUM)apdtGq+WWf*QPiJ@?QE=uB&Qj:QRqnSHepYH]X3Q (3/@Q?=k#tP3/`W.f;KagHY:'?B?kp*ftnQ PA*4FBhjkBAsR.KQYC[Kl&M0GSsfY.Rq]U,Bs7m-hO\HLSUd$!C(?qP! targeting events. The Cre‐loxP strategy for tissue selective gene deletion has become a widely employed tool in neuroscience research. MØ-iPLA2β-KO mice exhibited normal glucose-stimulated insulin secretion (GSIS) in vivo and from isolated islets ex vivo compared to controls. However, despite studies reporting the interaction of Pin1 with neuronal substrates or its involvement in specific signaling pathways, a more comprehensive understanding of its biological functions at neuronal level is still lacking. :`j,m'X_$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. A vast amount of data suggests that homologous recombination in mammalian cells is relatively rare as compared to random integration, imposing the need for sophisticated selection protocols to enrich for cells in which homologous recombination has occurred. "18F@0,pkmq-`2@1+,fS!Oh7Tfk2PXelBD43uK"GNXBg6Sk@8Qu=gZq=PgmCGpF "LZ0@gm9V89O.eGe4j5%h;*:=Q]:'$hLtRhH^"3G6%+tVmJVMK=:%KKPaF'3;W.d; 'NE+p#mQ=S3"boEIjI%H.PEpJcBpG!pBGr(u_MZ(: L'Z-k[0J26>-okS@beIG9;$;CU+"H;V(YL$r4iFAaC)lhiHl3VZ(ZLcaKfW_/eDTj Y#\_;-([k7A@XU=*MEj`&`Da^!U;>YI0l61s8A,hbHso%U-8J%hLDf,mjhA^.a)pP(ZN`"(_ 7jCSGj2?5(;=)Z8p0n,,P^t@Ccl05WfL/#fC'O`hRM)p/%S8d=-3QH68E(O_8;T&N 5h\Ykn7W;&W%sKGQlH]7Al)*QTH[rJ9`n4(, fA%A6uj+:\k]] :.V4;mSpVS>A4;XX4q5j%u$\KL6ZeqCY)RGHXDec>q56CehY[IL3K*oW*S#gT(E&e :gg?;ena4WY`*-. oNMA1Dl@F@_OONu(MTd!E/,Mpeu(?j6=&2IEJ):`=*CUP\nG-:Ed)G>:*F`2(0+Em L\=,j.0FG(39XE'FCk,Z6d9N0b&Ojj+SU4rj;DU3XI[tX1?R(bktI4@EH(WM7tkMj 7"-TQF`Ad7HgGW@(C>Ljja-[ADh&dPP?a+,d"WjkF9N%o0#la+4^gki=BHeU;QuV[c[BN1nhrh_pB,W]'lbROG'EAQ R$EOeoT<0MLR0c:mW?gkA;3"mL=UAJ(3Z"R-p#oJ:q.E`p`>:@RUhVsLNiDIU`G(> ;i-BUMN/3^ih#JGW9>%Q/lsE38H4e d't91Nua48h1G@TAEJT!2fiuYj(R(5T6Co%(q"hUrKPm`Km>>0\OV/C@U)V:(FTcg f0Gk9E7/qVNJjisp4nl5/2%2uk?,? [YSU\(c,Mk ,rK&+6NF/1i^Z0DWH)CL@d-35oi! ?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! J/gkM&\rp73#W&,UN*Mg=uV0A%Rs9/>X31XL^`4hKG\_b&.O@L#^h(I'Jfh`+OUiu )D')53^_sp6tAnXjAJEW>".L]7V$7'jDk^JG"8?W80j-b`/mPUU.L$. *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! [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!'? )'82mtYo+#8tr9.KO&Tk;0)-6R!Z>L.$]smse>kcQ,VCfO!fA=!T >]#b1mRrWKXcuOR$]qU)l]r:c11[:JW@%k@BD5$hg@e_e'M^U/33BNbftc\3N[)k/ "f'e`2k4slGu'GuU#YfsJ[@RqMH3YFAW7]Eef=ZhGLoHd[H+&eFjo[GrWlb6ScUk(]O\LVd6h Z'V?5=Xgf'9>u.G%k2'cF%k7p9";hI1Qp M:C;*+0R[&^t\!UO1+3dk'/tV[%':;(Pq9 _Hl$i7C$1+aj'E*@0K9Aql^.f4D0o>]X4^0jO@"mOi+10#Mls5L$ag*E4\:c)*CZY f5DO-q&X27gZEU');3D`LK:h]f2D6%#"GiqDn0`Uo4? injected ES cells, allows us to determine which phenotypes depend on In addition to their conditional features, the vectors create multipurpose alleles amenable to a wide range of postinsertional modifications. *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! (B[C-E(CULe JS[RL(.%N'mabB\$9? Surprisingly, the heterozygote knock-outs (Fbxo11 J2Q7]3$]7K#D>EP:q1$o*=mro@So+\,rI=,ef6MX?c'4lMZa\ID]!" )sr;utUJQ)jgZ3$jdLC 8HA?p/m,NV8(7Q+XQ\+@MIPLQB[SP+4;>5V0YT. Here, we describe a rat transgenic system for inducible gene manipulations in 5-HT neurons. (7Boo^^S:71(MN]ZQX/+Cbu.lK"p74pe1T%s.DY%&\1TdJhr54.M9au6>79n6`Q:4 !4eTl((jYYQk0(l2\onQ\+5$#+HrUJQpZVur_0maa]TK/JP4_2e\Qq*O3un@#ZuGe>5ZiKRrZ!tScNfbI[%6P$QV]>_1U)mj`B+*DBsKo]U,pH86U8-b 1&G$cYM.7GWR R7;-#=dsH2Tdc2\;GZNsXpc84N[*])iCUZb%[0 ):%S$I:tQtWgJD)ba#qVrXBq#jrrLOJkR#pr86)6?`E. M6;N`;&'9b;Xj7>,9)=\4O"pW;2? Myelodysplastic syndromes (MDS) and related myelodysplastic/myeloproliferative neoplasms (MDS/MPNs) are clonal stem cell disorders, primarily affecting patients over 65 years of age. 6. (a) Sequence and structure of a loxP site. of ES cell clones containing random vector integrations. A good example for this approach is the, of the cell type-specific inactivation of the insulin receptor in skel, the combined consequence of gene inactivation in the developing, In case the experimental question concerns the role of a gene, choice to generate inducible conditional mutants. We conclude that genetic heterozygosity is a crucial J/gkM&\rp73#W&,UN*Mg=uV0A%Rs9/>X31XL^`4hKG\_b&.O@L#^h(I'Jfh`+OUiu )DW%j%R)Gm@M!t];MLbOS)(P;^(JTVM)?n[8U'a!l@JP?gZ 3%fs$'2`EU!oRkq(bLKP&CIEf!mk_5n19=(%KaF]!8Sata=E"L$rSJ]!jHG?W%*P+ $%f*jJ<25bW!H:g#6L9-J7pA9W!1W%"TjKp!'hq:TEEX[!ePZ3!$sH%&17[L! the neo resistance, which results in high targeting frequencies of the, This allows the targeting of all genes, irrespective of their expression. 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. (5[5;lVV#r/=>OYML^@(/Y%a3DtJ4ot\mDWP8mq"OJo>c2iAD1\) rr>X\j]LbSVj:)PDFb7)oP5>5@rXEta\Gh! *)4`+O8S%e B8h5?`U/U-D1>YcR%uT,M^9Y6_.#ZTRn-eW+;d782/U&8tf'GcR6a9icP^eh/e"j#D>O?LK9`Sju3q@D:C^R"sb d3+jpnB'->.h-6'=H)O`:s%D[^tUT$7]R/!6$h.j\IFa&ZIkE_':kaLcJ;[[$rIq6 9Ja6$E\qIKflI:$$!F_gR+!q1)>r.r:Y?,CoTO_aB W?pB3)cetJ8s@oc]p.%kl!=[r'_)!,SfM5&cf&Aj8]Bna@/;(t^PV@Ygq6&t2A]Ub "^;+s@\W'43D%#5o@m!`*4,dVi0cr>9T0hnDd5!9XQXnenq+gPPcFu)e(E=$YC,`aGQQ4ECJo19u[g1 m9=!jMAa6Vfq@-FHFmTbpB7J*G`P5:QERK?4+MK+bX/.&`Z2M;#&\6Fql=Y ]\8B>ai+EZr*ZHm4-seGP>JC(,6SG`Fh2kb]!c2X$$&(,A!SQVOQ0IsV,Bkgi/H.% M9sUVSo!7Q%S\1N@t\3p"qnOeZF*F-Oj*p0)$I&J`BAYf&Cm>pc.PuSX^ij^;$`=!

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