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. 2022 May 5;13:2469. doi: 10.1038/s41467-022-30196-9

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Three main combinatorial CRISPR systems evaluated: eight distinct combinations of alternative spCas9 tracrRNAs sequences (orange boxes represent libraries generated), enCas12a, and orthologous spCas9-saCas9 system. b Schematic representation of the CRISPR screen pipeline. Ten distinct libraries are screened in IPC298. spCas9-saCas9, enCas12a, VCR1-WCR3, and WCR2-WCR3 libraries are screened in PK1 and MELJUSO. c Schematic of sgRNA constant regions with indicated alterations compared to original spCas9 tracrRNA. Dark blue represents canonical sequence; light blue represents regions where insertions are introduced; orange represents base changes.