TY - JOUR
T1 - Rapid in vivo multiplexed editing (RIME) of the adult mouse liver
AU - Katsuda, Takeshi
AU - Cure, Hector
AU - Sussman, Jonathan
AU - Simeonov, Kamen P.
AU - Krapp, Christopher
AU - Arany, Zoltan
AU - Grompe, Markus
AU - Stanger, Ben Z.
N1 - Funding Information:
We thank Jeffrey Posey at Oregon Health & Science University for vectors and advice on viral preparation and members of the Stanger laboratory for comments and helpful suggestions. We thank Salina Yuan for assisting with ATAC‐seq library preparation and the Penn Electron Microscopy Resource Laboratory for help with electron microscopy. This work was supported by NIH R01 DK083355 (to B.Z.S. and M.G.) and the Biesecker Pediatric Liver Foundation.
Publisher Copyright:
© 2022 American Association for the Study of Liver Diseases.
PY - 2022
Y1 - 2022
N2 - Background and Aims: Assessing mammalian gene function in vivo has traditionally relied on manipulation of the mouse genome in embryonic stem cells or perizygotic embryos. These approaches are time-consuming and require extensive breeding when simultaneous mutations in multiple genes is desired. The aim of this study is to introduce a rapid in vivo multiplexed editing (RIME) method and provide proof of concept of this system. Approach and Results: RIME, a system wherein CRISPR/caspase 9 technology, paired with adeno-associated viruses (AAVs), permits the inactivation of one or more genes in the adult mouse liver. The method is quick, requiring as little as 1 month from conceptualization to knockout, and highly efficient, enabling editing in >95% of target cells. To highlight its use, we used this system to inactivate, alone or in combination, genes with functions spanning metabolism, mitosis, mitochondrial maintenance, and cell proliferation. Conclusions: RIME enables the rapid, efficient, and inexpensive analysis of multiple genes in the mouse liver in vivo.
AB - Background and Aims: Assessing mammalian gene function in vivo has traditionally relied on manipulation of the mouse genome in embryonic stem cells or perizygotic embryos. These approaches are time-consuming and require extensive breeding when simultaneous mutations in multiple genes is desired. The aim of this study is to introduce a rapid in vivo multiplexed editing (RIME) method and provide proof of concept of this system. Approach and Results: RIME, a system wherein CRISPR/caspase 9 technology, paired with adeno-associated viruses (AAVs), permits the inactivation of one or more genes in the adult mouse liver. The method is quick, requiring as little as 1 month from conceptualization to knockout, and highly efficient, enabling editing in >95% of target cells. To highlight its use, we used this system to inactivate, alone or in combination, genes with functions spanning metabolism, mitosis, mitochondrial maintenance, and cell proliferation. Conclusions: RIME enables the rapid, efficient, and inexpensive analysis of multiple genes in the mouse liver in vivo.
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U2 - 10.1002/hep.32759
DO - 10.1002/hep.32759
M3 - Article
C2 - 36037289
AN - SCOPUS:85139663149
SN - 0270-9139
JO - Hepatology
JF - Hepatology
ER -