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

Autor:	Li S; Department of Molecular Biology, Princeton University, Princeton, NJ 08540, USA., Mereby SA; Department of Molecular Biology, Princeton University, Princeton, NJ 08540, USA., Rothstein M; Department of Molecular Biology, Princeton University, Princeton, NJ 08540, USA., Johnson MR; Department of Molecular Biology, Princeton University, Princeton, NJ 08540, USA., Brack BJ; Department of Molecular Biology, Princeton University, Princeton, NJ 08540, USA., Mallarino R; Department of Molecular Biology, Princeton University, Princeton, NJ 08540, USA. Electronic address: rmallarino@princeton.edu.
Jazyk:	angličtina
Zdroj:	Cell reports [Cell Rep] 2023 Aug 29; Vol. 42 (8), pp. 112980. Date of Electronic Publication: 2023 Aug 11.
DOI:	10.1016/j.celrep.2023.112980
Abstrakt:	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. Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu