Strain Development
CRISPR RNP was used to assist with homologous recombination using dsDNA repair template in JM8A3 ES cells derived from C57BL/6N for generation of the Ace2 deletion and ACE2 insertion model . Following blastocyst injection, the chimeras were mated with C57BL/6NCrl to create germline heterozygotes. Long-range PCR products were fully sequenced; potential off-targets were absent. CRISPR/RNP was used to assist with homologous recombination using a dsDNA repair template directly in zygotes for the Dpp4 deletion and DPP4 insertion model. The founder was backcrossed with C57BL/6NCrl to create germline heterozygous animals. Long-range PCR products were sequenced in-full to confirm the absence of potential off-targets. One additional backcross to C57BL/6NCrl. The two targeted lines were then intercrossed to derive mice with both mutations. Mice are homozygous (females) or hemizygous (males) for humanized ACE2, and homozygous for humanized DPP4.
Suggested Control Mice
Sibling wild types or C57BL/6N