Protocol for in vivo CRISPR screening using selective CRISPR antigen removal lentiviral vectors
Sarah Kate Lane-Reticker 1, Emily A Kessler 1, Audrey J Muscato 1, Sarah Y Kim 1, John G Doench 1, Kathleen B Yates 2, Robert T Manguso 3, Juan Dubrot 4
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 the Renca mouse model. This protocol describes conducting 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.
Keywords: CRISPR; Cancer; Immunology; Sequencing.