TIGER: Single-step in vivo genome editing in a non-traditional rodent

Sha Li, Sarah A. Mereby, Megan Rothstein, Matthew R. Johnson, Benjamin J. Brack, Ricardo Mallarino

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Article number112980
JournalCell Reports
Volume42
Issue number8
DOIs
StatePublished - Aug 29 2023

All Science Journal Classification (ASJC) codes

  • General Biochemistry, Genetics and Molecular Biology

Keywords

  • African striped mice
  • CP: Developmental biology
  • CP: Molecular biology
  • CRISPR
  • CRISPR knock-in
  • CRISPR knock-out
  • Rhabdomys pumilio
  • emerging model organisms
  • in vivo genome editing
  • intraoviductal injections
  • non-traditional rodents

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