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. 2021 Jul 9;50(5):e25. doi: 10.1093/nar/gkab519

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — CRISPRpas alters intronic APA of endogenous genes. (A) UCSC tracks of human RAD51C gene structure and 3′READS+ data for FT and 4sU fractions (top). The three APA sites and Stability Scores are indicated (bottom). A gRNA was designed to target a region in intron 2. Primers for detection of IPA isoform and last exon PAS isoforms are indicated. (B) RT-qPCR analysis of relative amounts of last exon PAS isoforms (blue) and IPA isoform (orange) in HEK293T^dCas9 cells transfected with target or Ctrl gRNAs (48 h). The ratio of IPA isoform/last exon PAS isoform is also shown (gray). Data were normalized to Ctrl gRNA. Error bars are from standard error of mean (n = 3). P-value is calculated from student's t-test. n.s., not significant; *,P < 0.05; **, P < 0.01; ***, P < 0.001. (C) As in (A), except that human ANKMY1 gene structure and data are shown. A gRNA was designed to target a region in intron 7. (D) As in (B), except that data for ANKMY1 are shown. Error bars are standard error of means (n = 4).