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. 2022 Feb 22;50(5):e26. doi: 10.1093/nar/gkac108

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© The Author(s) 2022. 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 3. — dPguCas13b enabled both repression and enhancement of the intronic PAS usage. (A) Illustration of genes with intronic APA. The positions of gRNA target regions were indicated. (B) UCSC genome browser tracks for RNA-Seq and 3′end mRNA Seq were shown for the human gene AK2. The intronic PAS with the dominant usage (Intronic PAS) and the distal PAS (disPAS) were indicated. (C) The usage of the intronic PAS in gene AK2 was repressed after transfection of 3xEGFP-dPguCas13b together with gRNAs targeting the PAS signal (gPAS) and USE (gUSE), but enhanced with gRNA targeting the 3′ splicing site (g3′SS). (D) UCSC genome browser tracks for RNA-Seq and 3′end mRNA Seq was shown for the human gene PCMT1. The intronic PAS with the dominant usage (Intronic PAS) and the distal PAS (disPAS) were indicated. (E) The usage of the intronic PAS in gene PCMT1 was repressed after transfection of 3xEGFP-dPguCas13b together with gRNAs targeting the PAS signal (gPAS) and USE (gUSE), but enhanced with gRNA targeting the 3′ splicing site (g3′SS). For each experiment, three independent repeats were performed. Error bars represent SEM. ****P < 0.0001, paired two-way Student's t-test.