TIGER: Single-step in vivo genome editing in a non-traditional rodent. Author Sha Li, Sarah Mereby, Megan Rothstein, Matthew Johnson, Benjamin Brack, Ricardo Mallarino Publication Year 2023 Type Journal Article Abstract Rodents are taxonomically diverse and have evolved a variety of traits. A mechanistic understanding of such traits has remained elusive, however, largely because genome editing in non-traditional model species remains challenging. Here, using the African striped mouse (Rhabdomys pumilio), we describe TIGER (targeted in vivo genome editing in rodents), a method that relies on a simple intraoviductal injecting technique and uses recombinant adeno-associated viruses (rAAVs) as the sole vehicle to deliver reagents into pregnant females. We demonstrate that TIGER generates knockout and knockin (up to 3 kb) lines with high efficiency. Moreover, we engineer a double-cleaving repair rAAV template and find that it significantly increases knockin frequency and germline transmission rates. Lastly, we show that an oversized double-cleaving rAAV template leads to an insertion of 3.8 kb. Thus, TIGER constitutes an attractive alternative to traditional ex vivo genome-editing methods and has the potential to be extended to a broad range of species. Keywords African striped mice, CP: Developmental biology, CP: Molecular biology, CRISPR, CRISPR knock-in, CRISPR knock-out, Rhabdomys pumilio, emerging model organisms, intraoviductal injections, in vivo genome editing, non-traditional rodents Journal Cell reports Volume 42 Issue 8 Pages 112980 Date Published 08/2023 ISSN Number 2211-1247 DOI 10.1016/j.celrep.2023.112980 Alternate Journal Cell Rep PMCID PMC10528174 PMID 37573509 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML