Hprt1^{tm1(Gdf9-cas9)Jahe}

ES cell line mEMS6131.02 (C57BL/6NTac-A(w-J)/A(w-J), Hprt1(b-m3)/Y; J:270567) was targeted with Hprt targeting vector pEMS-2001 (J:144244) harboring a 3.3 fragment of the mouse Gdf9 promoter from pGDF-9-iCre (J:93180) and S. pyogenes Cas9 from pX330-U6-Chimeric_BB-CBh-hSpCas9 (PMID: 23287718). HAT medium was used for selection and properly targeted ES cell clones were injected into C57BL/6NTac-Tyr(tm1Arte) blastocysts. Resulting chimeras were bred to C57BL/6NJ animals for germline transmission. The Gdf9 promoter drives Cas9 expression in oocytes starting at the primordial ovarian follicle stage. Maternally (oocyte) supplied S.p. Cas9 facilitates genome editing in zygotes electroporated or microinjected with a guide RNA. (J:337011)