Generation Of Gata-1 Tissue-specific Knockout Mouse Model Using CRISPR/Cas9 System

The transcription factor Gata1 is essential for the development of erythroid cells.Consequently,Gata1 null mutants die in utero due to severe anemia.Outside the hematopoietic system,Gata1 was thought to be only expressed in the Sertoli cells of the testis.But our lab has recently demonstrated that GATA-1 is also expressed in adipose tissue.To elucidate the function of Gata1 in the development of adipose tissue,we made an adipose tissue-specific knockout of the Gata1 gene in the mouse using CRISPR/Cas9 system.The CRISPR/Cas system has been adapted as an efficient gene-targeting technology with the potential for multiplexed genome editing.During the present study,we used 4 sg RNAs(Sg2,Sg3,Sg4,Sg5)targeting human GATA-1 and 3 Sg RNAs(Sg1,Sg2,Sg3,)targeting mouse GATA-1 gene.Upon validation,only Sg RNAs working at cell level were used to generate adipose tissuespecific Knockout mice under ap2 promoter.The transgenic mice were crossed among them for germline transmission before sacrificing them.The study is still in process We show that this process which relies on CRISPR components is sequence-specific and,upon simultaneous introduction of multiple g RNAs,can effect multiplex editing of target loci.We also compute a 606 bps and more than 1Kb deletions respectively in mouse and human GATA-1 genes.Our results establish an RNA-guided editing tool for facile,robust,and multiplexable genome engineering.