Plscr_Q271Y was created in the mouse to model the human coding variant PLSCR_H262Y caused by rs343320. CRISPR RNP was used to assist with homology directed repair (HDR) using an ssODN repair template electroporated in mouse C57BL/6N-Ace2em1(ACE2)Mbp Tmprss2em1(TMPRSS2)Mbp zygotes. The targeted residue codon CAG to TAT at the 271th residue were engineered into the HDR template totaling 2 guide mismatches to prevent RNP re-cleavage. Founder animals were screened by QPCR MGB probe with positives subsequently PCR’d and bidirectionally sequenced at the loci.

Not applicable

Unknown

Plscr_Q271Y was created in the mouse to model the human coding variant PLSCR_H262Y caused by rs343320. CRISPR RNP was used to assist with homology directed repair (HDR) using an ssODN repair template electroporated in mouse C57BL/6N-Ace2em1(ACE2)Mbp Tmprss2em1(TMPRSS2)Mbp zygotes. The targeted residue codon CAG to TAT at the 271th residue were engineered into the HDR template totaling 2 guide mismatches to prevent RNP re-cleavage. Founder animals were screened by QPCR MGB probe with positives subsequently PCR’d and bidirectionally sequenced at the loci. Sequence confirmed founder animals were then backcrossed to C57BL/6N-Ace2em1(ACE2)Mbp Tmprss2em1(TMPRSS2)Mbp and subsequence N1 heterozygous animals we confirmed for copy number as well as sequence analysis. Potential off targets with an MIT score greater than 1.5 were not found.

C57BL/6NCrl or wild-type siblings

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.

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