ࡱ> 241[(bjbj .ΐΐ||  7$/-ZZZ@ZZZZ]P@Z07ZF ZZ0ZP 7| :Generation of mice with inducible T-cell specific expression of Cre Antulov Ronald, Mandari Sanja, Zafirova Biljana, Arapovi Jurica, Poli Bojan Department of Histology and Embryology, University of Rijeka School of Medicine, Croatia Inducible Cre/lox P system represents an effective tool to study the role of specific gene in vivo, providing spatial (cell type specific) and temporal (inducible) control of gene expression at the same time. Here, we report the generation and characterization of a mouse mutant strain with inducible T cell-specific control of Cre, which we have developed to study the role of several genes in the activation and homeostasis of mature T cells in vivo. We used inducible CreER T2 system where the activity of CreER fusion protein can be induced by administration of tamoxifen. T cell specificity and inducibility of Cre were provided by knocking of the CreERT2 cassette into the CD3e locus. We designed an appropriate targeting vector that was electroporated into the Bruce4 (C57BL6) embryonic stem (ES) cells. Upon the selection and screening procedure, we identified several ES cell clones positive for the homologous recombination. Selected ES clones were microinjected into mouse blastocysts and several chimeras were obtained that gave the germline transmission of the mutation. We crossed Cd3e-CreERT2 mice to TCR Cafl mice, containing loxP flanked TCR Ca region, in order to examine the efficiency of tamoxifen inducible Cre mediated deletion in T cells. In this system, we showed conditional TCRab ablation on about 30% of CD4+ and CD8+ T cells mediated by Cre recombination upon tamoxifen administration. The specificity of Cre expression and quantification of Cre mediated deletion in different cell types and tissues were d %&(hkXUh}h(&Jh}hH_5h}h}5 & &'($a$gd(&Jgd(&J etermined by quantitative PCR method. ,1h. A!"#$% ^ 2 0@P`p2( 0@P`p 0@P`p 0@P`p 0@P`p 0@P`p 0@P`p8XV~_HmHnHsHtH@`@ NormalCJ_HaJmHsHtHDA D Default Paragraph FontRiR  Table Normal4 l4a (k (No List PK![Content_Types].xmlj0Eжr(΢Iw},-j4 wP-t#bΙ{UTU^hd}㨫)*1P' ^W0)T9<l#$yi};~@(Hu* Dנz/0ǰ $ X3aZ,D0j~3߶b~i>3\`?/[G\!-Rk.sԻ..a濭?PK!֧6 _rels/.relsj0 }Q%v/C/}(h"O = C?hv=Ʌ%[xp{۵_Pѣ<1H0ORBdJE4b$q_6LR7`0̞O,En7Lib/SeеPK!kytheme/theme/themeManager.xml M @}w7c(EbˮCAǠҟ7՛K Y, e.|,H,lxɴIsQ}#Ր ֵ+!,^$j=GW)E+& 8PK!Ptheme/theme/theme1.xmlYOo6w toc'vuر-MniP@I}úama[إ4:lЯGRX^6؊>$ !)O^rC$y@/yH*񄴽)޵߻UDb`}"qۋJחX^)I`nEp)liV[]1M<OP6r=zgbIguSebORD۫qu gZo~ٺlAplxpT0+[}`jzAV2Fi@qv֬5\|ʜ̭NleXdsjcs7f W+Ն7`g ȘJj|h(KD- dXiJ؇(x$( :;˹! I_TS 1?E??ZBΪmU/?~xY'y5g&΋/ɋ>GMGeD3Vq%'#q$8K)fw9:ĵ x}rxwr:\TZaG*y8IjbRc|XŻǿI u3KGnD1NIBs RuK>V.EL+M2#'fi ~V vl{u8zH *:(W☕ ~JTe\O*tHGHY}KNP*ݾ˦TѼ9/#A7qZ$*c?qUnwN%Oi4 =3ڗP 1Pm \\9Mؓ2aD];Yt\[x]}Wr|]g- eW )6-rCSj id DЇAΜIqbJ#x꺃 6k#ASh&ʌt(Q%p%m&]caSl=X\P1Mh9MVdDAaVB[݈fJíP|8 քAV^f Hn- "d>znNJ ة>b&2vKyϼD:,AGm\nziÙ.uχYC6OMf3or$5NHT[XF64T,ќM0E)`#5XY`פ;%1U٥m;R>QD DcpU'&LE/pm%]8firS4d 7y\`JnίI R3U~7+׸#m qBiDi*L69mY&iHE=(K&N!V.KeLDĕ{D vEꦚdeNƟe(MN9ߜR6&3(a/DUz<{ˊYȳV)9Z[4^n5!J?Q3eBoCM m<.vpIYfZY_p[=al-Y}Nc͙ŋ4vfavl'SA8|*u{-ߟ0%M07%<ҍPK! ѐ'theme/theme/_rels/themeManager.xml.relsM 0wooӺ&݈Э5 6?$Q ,.aic21h:qm@RN;d`o7gK(M&$R(.1r'JЊT8V"AȻHu}|$b{P8g/]QAsم(#L[PK-![Content_Types].xmlPK-!֧6 +_rels/.relsPK-!kytheme/theme/themeManager.xmlPK-!Ptheme/theme/theme1.xmlPK-! ѐ' theme/theme/_rels/themeManager.xml.relsPK] (( 8@0(  B S  ?CDCDPp(&JkXH_}]!@(@(@UnknownG* Times New Roman5Symbol3. * ArialACambria Math"7ˆ7ˆ  !r42HX)?}2!xxCGeneration of mice with inducible T-cell specific expression of Cre korisnik1biljanaOh+'0, <H h t DGeneration of mice with inducible T-cell specific expression of Cre korisnik1Normalbiljana2Microsoft Office Word@@"P@"P՜.+,0H hp  ( Medicinski fakultet u Rijeci  DGeneration of mice with inducible T-cell specific expression of Cre Title  "#$%&'(*+,-./03Root Entry Fp(P5Data  1TableWordDocument.SummaryInformation(!DocumentSummaryInformation8)CompObjy  F'Microsoft Office Word 97-2003 Document MSWordDocWord.Document.89q