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

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 genomeediting methods and has the potential to be extended to a broad range of species.

Automated summary

The study presents a method called TIGER, which utilizes intraoviductal injecting technique and recombinant adeno-associated viruses (rAAVs) to deliver reagents into pregnant females for genome editing in non-traditional model species. The method shows high efficiency in generating knockout and knockin lines, and the use of a double-cleaving repair rAAV template significantly increases knockin frequency and germline transmission rates. The study suggests that TIGER could be a promising alternative to ex vivo genome editing methods and can be applied to a wide range of species.