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

Sarah Kate Lane-Reticker; Emily A. Kessler; Audrey J. Muscato; Sarah Y. Kim; John G. Doench; Kathleen B. Yates; Robert T. Manguso; Juan Dubrot

STAR Protocols (Mar 2023)

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

Sarah Kate Lane-Reticker,
Emily A. Kessler,
Audrey J. Muscato,
Sarah Y. Kim,
John G. Doench,
Kathleen B. Yates,
Robert T. Manguso,
Juan Dubrot

Affiliations

Sarah Kate Lane-Reticker: Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA
Emily A. Kessler: Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA
Audrey J. Muscato: Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA
Sarah Y. Kim: Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA
John G. Doench: Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA
Kathleen B. Yates: Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Center for Cancer Research, Massachusetts General Hospital, Boston, MA 02114, USA
Robert T. Manguso: Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Center for Cancer Research, Massachusetts General Hospital, Boston, MA 02114, USA; Corresponding author
Juan Dubrot: 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; Corresponding author

Journal volume & issue: Vol. 4, no. 1
p. 102082

Abstract

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Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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