Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 Jan 21;13:437. doi: 10.1038/s41467-022-28135-9

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 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/.

PMC Copyright notice

Fig. 3 — a Comparison of off-target site detection for a promiscuous sgRNA targeting Pcsk9 (Pcsk9-gP or sgPcsk9) by GUIDE-tag, VIVO, and Discover-seq. For GUIDE-tag analysis genomic DNA was isolated from the liver of mice injected with plasmid expressing sgPcsk9-SpyCas9-mSA* plus iGUIDE or biotin-iGUIDE donors (n = 3). UMI numbers recovered for each site are shown (Average of three mice). Mismatches relative to the on-target site are shown with colored boxes. Indel frequencies determined by targeted amplicon sequencing from the liver of three mice are presented as a heat map (1,2,3 are three mice) and average indel% (right). Forty of 52 tested GUIDE-tag discovered off-target sites show statistically significant indels (p < 0.05) (ranked by average indel%). N.V, not verified, identified by CIRCLE-seq but not chosen for validation in the VIVO study. Dash, off-target site not identified by VIVO or DISCOVER-seq. b Venn diagram of the overlap of validated off-target cleavage sites discovered by GUIDE-tag (40 validated loci), VIVO (19 validated loci), and DISCOVER-seq (26 loci) for sgPcsk9. Note only a subset of potential off-target sites were tested in each study. c Scatter plots of average indel frequency (x-axis, targeted amplicon deep sequencing) and average UMI% (y-axis, n = 3 mice) for verified sgPcsk9 off-target sites identified by GUIDE-tag. Dashed lines represent the linear regression fit (Spearman’s correlation calculated). The p-value for Pearson’s correlation coefficient was determined by the two-tailed t-distribution table. Source data are provided as a Source Data file.