Transcriptome signature of miRNA-26b KO mouse model suggests novel transcriptional interactions
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https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE147519
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miR26b KO mice were generated by Laser assisted (XY-Clone Hamilton Thorne) injection of R1/E(129/Sv) cells into 8-cell stage C57Bl/6NCrl embryos.Chimaeras were crossed to C57Bl/6NCrl mice and their offspring was screened for germline transmission. Embryo donor and recipient mice: C57BL/6NCrl, Crl:CD1(ICR). We induced homologous recombination in embryonic stem (ES) cells using a construct containing 2.0 kb of gDNA, including exon 2,3 and 4, as also part of intron 4-5, a Neo Cassette was inserted in between Frt sites. 160 bp of gDNA, including miR26b was flanked by loxP sites (floxed). The construct was finished by introducing a 5.6 kb fragment of gDNA serving as 3’ recombination arm.A correctly targeted ES cell clones were injected into blastocysts to produce a chimeric mice which transmitted the modified allele through the germ line. A male heterozygous for the targeted allele was bred with a female expressing ubiquitous Flippase (Flp) transgene to ultimately produce animals that had deleted the Neo cassette, preserving the loxP sites flanking exon miR26b. Compare differential gene expression between naïve mouse and miRNA26B K.O
创建时间:
2021-07-14



