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Rapid in vivo multiplexed editing (RIME) of the adult mouse liver
Takeshi Katsuda1,2,3, Hector Cure1,2,3, Jonathan Sussman1,2,3
1Department of Medicine , University of Pennsylvania , Philadelphia , Pennsylvania , USA.
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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