Comparative analysis of single-stranded DNA donors to generate conditional null mouse alleles.

ABSTRACT:

Background

The International Mouse Phenotyping Consortium is generating null allele mice for every protein-coding gene in the genome and characterizing these mice to identify gene-phenotype associations. While CRISPR/Cas9-mediated null allele production in mice is highly efficient, generation of conditional alleles has proven to be more difficult. To test the feasibility of using CRISPR/Cas9 gene editing to generate conditional knockout mice for this large-scale resource, we employed Cas9-initiated homology-driven repair (HDR) with short and long single stranded oligodeoxynucleotides (ssODNs and lssDNAs).

Results

Using pairs of single guide RNAs and short ssODNs to introduce loxP sites around a critical exon or exons, we obtained putative conditional allele founder mice, har

SUBMITTER: Lanza DG 

PROVIDER: S-EPMC6011517 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature