The mouse gene was deleted and the human gene simultaneously inserted via CRISPR targeting for each, Ace2 and Dpp4, in separate targeting events.

JM8A3 derived from C57BL/6N

Unknown

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.

Sibling wild types or C57BL/6N

The MMRRC Centers have developed a genetic QC pipeline using MiniMUGA array genotyping to provide additional information on strain backgrounds for MMRRC congenic and inbred strains. For more information on when data may be available, or to request genotyping for a strain of interest, please contact mmrrc@ucdavis.edu. Older strains may not have this information.

Kent Lloyd, D.V.M., University of California, Davis.

Not ready to publish