TABLE 1.
Summary of current tools to alter PAS usagea
| Category | Method | Mode of action | Permanent PAS change | Amenable to HTS | Representative studies |
|---|---|---|---|---|---|
| Masking of PAS motifs | ASO | Masking PAS motif; inhibiting interaction between PAS and CPA machinery | No | No | Chen et al. 2016 |
| U7 SmOPT | Antisense sequence in engineered U7-snRNA; masking PAS motif | No | No | Rashnonejad et al. 2021 | |
| CRISPR/dCas13 | Masking PAS motif by dCas13-gRNA | No | Yes | Tian et al. 2022 | |
| Genomic DNA editing | CRISPR/Cas9 + a pair of gRNAs flanking PAS | Removal of PAS from the genome; DNA repair by NHEJ | Yes | Yes | Kamieniarz-Gdula et al. 2019; Wang et al. 2019; Gabel et al. 2024. |
| CRISPR/Cas9 + a pair of gRNAs + donor sequence | PAS removal or motif change through double-stranded DNA cut and HDR | Yes | No | Wang et al. 2018a; Bae et al. 2020 | |
| CRISPR/adenine base editor (ABE) | Genomic base editing of PAS motif | Yes | Yes | Sikrova et al. 2021 | |
| RNAPII elongation block | CRISPR/dCas9 | Blocking RNAPII elongation by dCas9 to promote alternative PAS usage | No | Yes | Shin et al. 2022; Zukher et al. 2023 |
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(HTS) High-throughput screening.
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aIndirect PAS usage alteration through splicing is not included.
