Programmed genome editing by a miniature CRISPR-Cas12f nuclease - PubMed (original) (raw)
. 2021 Nov;17(11):1132-1138.
doi: 10.1038/s41589-021-00868-6. Epub 2021 Sep 2.
Affiliations
- PMID: 34475565
- DOI: 10.1038/s41589-021-00868-6
Programmed genome editing by a miniature CRISPR-Cas12f nuclease
Zhaowei Wu et al. Nat Chem Biol. 2021 Nov.
Abstract
The RNA-guided CRISPR-associated (Cas) nucleases are versatile tools for genome editing in various organisms. The large sizes of the commonly used Cas9 and Cas12a nucleases restrict their flexibility in therapeutic applications that use the cargo-size-limited adeno-associated virus delivery vehicle. More compact systems would thus offer more therapeutic options and functionality for this field. Here, we report a miniature class 2 type V-F CRISPR-Cas genome-editing system from Acidibacillus sulfuroxidans (AsCas12f1, 422 amino acids). AsCas12f1 is an RNA-guided endonuclease that recognizes 5' T-rich protospacer adjacent motifs and creates staggered double-stranded breaks to target DNA. We show that AsCas12f1 functions as an effective genome-editing tool in both bacteria and human cells using various delivery methods, including plasmid, ribonucleoprotein and adeno-associated virus. The small size of AsCas12f1 offers advantages for cellular delivery, and characterizations of AsCas12f1 may facilitate engineering more compact genome-manipulation technologies.
© 2021. The Author(s), under exclusive licence to Springer Nature America, Inc.
Similar articles
- CRISPR-AsCas12f1 couples out-of-protospacer DNA unwinding with exonuclease activity in the sequential target cleavage.
Song X, Chen Z, Sun W, Yang H, Guo L, Zhao Y, Li Y, Ren Z, Shi J, Liu C, Ma P, Huang X, Ji Q, Sun B. Song X, et al. Nucleic Acids Res. 2024 Dec 11;52(22):14030-14042. doi: 10.1093/nar/gkae989. Nucleic Acids Res. 2024. PMID: 39530229 Free PMC article. - Assessment of Miniature AsCas12f1 Variants for Gene Editing and Activation.
Ren C, Bao Z. Ren C, et al. Biotechnol Bioeng. 2025 Jun;122(6):1590-1597. doi: 10.1002/bit.28978. Epub 2025 Mar 19. Biotechnol Bioeng. 2025. PMID: 40108777 - Miniature type V-F CRISPR-Cas nucleases enable targeted DNA modification in cells.
Bigelyte G, Young JK, Karvelis T, Budre K, Zedaveinyte R, Djukanovic V, Van Ginkel E, Paulraj S, Gasior S, Jones S, Feigenbutz L, Clair GS, Barone P, Bohn J, Acharya A, Zastrow-Hayes G, Henkel-Heinecke S, Silanskas A, Seidel R, Siksnys V. Bigelyte G, et al. Nat Commun. 2021 Oct 26;12(1):6191. doi: 10.1038/s41467-021-26469-4. Nat Commun. 2021. PMID: 34702830 Free PMC article. - Efficient Genome Editing by a Miniature CRISPR-AsCas12f1 Nuclease in Bacillus anthracis.
Wang Y, Sang S, Zhang X, Tao H, Guan Q, Liu C. Wang Y, et al. Front Bioeng Biotechnol. 2022 Jan 14;9:825493. doi: 10.3389/fbioe.2021.825493. eCollection 2021. Front Bioeng Biotechnol. 2022. PMID: 35096801 Free PMC article. Review. - Miniature CRISPR-Cas12 Systems: Mechanisms, Engineering, and Genome Editing Applications.
Tang N, Ji Q. Tang N, et al. ACS Chem Biol. 2024 Jul 19;19(7):1399-1408. doi: 10.1021/acschembio.4c00247. Epub 2024 Jun 20. ACS Chem Biol. 2024. PMID: 38899980 Review.
Cited by
- PAM-flexible Engineered FnCas9 variants for robust and ultra-precise genome editing and diagnostics.
Acharya S, Ansari AH, Kumar Das P, Hirano S, Aich M, Rauthan R, Mahato S, Maddileti S, Sarkar S, Kumar M, Phutela R, Gulati S, Rahman A, Goel A, Afzal C, Paul D, Agrawal T, Pulimamidi VK, Jalali S, Nishimasu H, Mariappan I, Nureki O, Maiti S, Chakraborty D. Acharya S, et al. Nat Commun. 2024 Jun 28;15(1):5471. doi: 10.1038/s41467-024-49233-w. Nat Commun. 2024. PMID: 38942756 Free PMC article. - The application of CRISPR/Cas technologies to Brassica crops: current progress and future perspectives.
Li J, Yu X, Zhang C, Li N, Zhao J. Li J, et al. aBIOTECH. 2022 Jul 2;3(2):146-161. doi: 10.1007/s42994-022-00076-3. eCollection 2022 Jun. aBIOTECH. 2022. PMID: 36304520 Free PMC article. Review. - Efficient genome editing in rice with miniature Cas12f variants.
Ye Z, Zhang Y, He S, Li S, Luo L, Zhou Y, Tan J, Wan J. Ye Z, et al. aBIOTECH. 2024 May 28;5(2):184-188. doi: 10.1007/s42994-024-00168-2. eCollection 2024 Jun. aBIOTECH. 2024. PMID: 38974870 Free PMC article. - Miniature CRISPR-Cas12f1-Mediated Single-Nucleotide Microbial Genome Editing Using 3'-Truncated sgRNA.
Lee HJ, Kim HJ, Lee SJ. Lee HJ, et al. CRISPR J. 2023 Feb;6(1):52-61. doi: 10.1089/crispr.2022.0071. Epub 2022 Dec 23. CRISPR J. 2023. PMID: 36576897 Free PMC article. - Development of CRISPR Cas9, spin-off technologies and their application in model construction and potential therapeutic methods of Parkinson's disease.
Qu J, Liu N, Gao L, Hu J, Sun M, Yu D. Qu J, et al. Front Neurosci. 2023 Jul 6;17:1223747. doi: 10.3389/fnins.2023.1223747. eCollection 2023. Front Neurosci. 2023. PMID: 37483347 Free PMC article. Review.
References
- Makarova, K. S. et al. Evolutionary classification of CRISPR-Cas systems: a burst of class 2 and derived variants. Nat. Rev. Microbiol. 18, 67–83 (2020). - DOI
- Jinek, M. et al. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337, 816–821 (2012). - DOI
- Cong, L. et al. Multiplex genome engineering using CRISPR/Cas systems. Science 339, 819–823 (2013). - DOI
- Mali, P. et al. RNA-guided human genome engineering via Cas9. Science 339, 823–826 (2013). - DOI
- Zetsche, B. et al. Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell 163, 759–771 (2015). - DOI
Publication types
MeSH terms
Substances
Supplementary concepts
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials