Protocol for in vivo CRISPR screening using selective CRISPR antigen removal lentiviral vectors.

Autor:	Lane-Reticker SK; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Kessler EA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Muscato AJ; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Kim SY; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Doench JG; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Yates KB; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Center for Cancer Research, Massachusetts General Hospital, Boston, MA 02114, USA., Manguso RT; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Center for Cancer Research, Massachusetts General Hospital, Boston, MA 02114, USA. Electronic address: rmanguso@broadinstitute.org., Dubrot J; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Solid Tumors Program, Division of Oncology, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain. Electronic address: jdubrot@unav.es.
Jazyk:	angličtina
Zdroj:	STAR protocols [STAR Protoc] 2023 Mar 17; Vol. 4 (1), pp. 102082. Date of Electronic Publication: 2023 Feb 01.
DOI:	10.1016/j.xpro.2023.102082
Abstrakt:	Recognition of Cas9 and other proteins encoded in delivery vectors has limited CRISPR technology in vivo. Here, we present a protocol for genome engineering using selective CRISPR antigen removal (SCAR) lentiviral vectors in Renca mouse model. This protocol describes how to conduct an in vivo genetic screen with a sgRNA library and SCAR vectors that can be applied to different cell lines and contexts. For complete details on the use and execution of this protocol, please refer to Dubrot et al. (2021). 1 . Competing Interests: Declaration of interests This work was supported in part by funding from Calico Life Sciences. J.D., J.G.D., K.B.Y., and R.T.M. are authors of a patent application related to the SCAR vector system. (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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