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. 2024 Jan 19;25:25. doi: 10.1186/s13059-023-03132-3

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — New optimizations in the cloning protocol improve genome-wide guide libraries. A Histograms of the CRISPRi V2 legacy library (blue) compared to the optimized CRISPRi V2 LGR library (orange) show that the LGR library has a tighter distribution of sgRNAs. B Similarly, the CRISPRa V2 LGR library (orange) shows a tighter distribution than the CRISPRa V2 legacy library (blue). C Skew ratio table comparing CRISPRi V2 and CRISPRa V2 LGR libraries to the legacy libraries. D Skew ratios of publicly available CRISPR libraries (Addgene catalog numbers available in Additional file 4) at the 90/10, 95/5, 98/2, 99/1, and 99.5/0.5 percentiles compared to the CRISPRi and CRISPRa LGR libraires