Miniature type V-F CRISPR-Cas nucleases enable targeted DNA modification in cells
| Autor: | Karolina Budre, Stephen L. Gasior, Selgar Henkel-Heinecke, Tautvydas Karvelis, Ralf Seidel, Virginijus Siksnys, Vesna Djukanovic, Rimante Zedaveinyte, Spencer Jones, Sushmitha Paulraj, Arunas Silanskas, Elizabeth Van Ginkel, Grace St. Clair, Greta Bigelyte, Pierluigi Barone, Gina Zastrow-Hayes, Ananta Acharya, Jennifer A. Bohn, Joshua K. Young, Lanie Feigenbutz |
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| Jazyk: | angličtina |
| Předmět: |
CRISPR-Cas9 genome editing
CRISPR-Cas targeted DNA modification Class 2 CRISPR systems Science CRISPR-Associated Proteins General Physics and Astronomy Molecular engineering in plants Zea mays Biochemistry Article General Biochemistry Genetics and Molecular Biology chemistry.chemical_compound Genome editing Plant Cells Cleave Humans CRISPR Gene Editing chemistry.chemical_classification Bacillales Clostridiales Nuclease Endodeoxyribonucleases Multidisciplinary biology Effector Cas9 DNA General Chemistry Cell biology Amino acid HEK293 Cells Ribonucleoproteins chemistry biology.protein CRISPR-Cas Systems Protein Multimerization RNA Guide Kinetoplastida |
| Zdroj: | Nature communications, Berlin : Nature research, 2021, vol. 12, iss. 1, art. no. 6191, p. [1-8] Nature Communications Nature Communications, Vol 12, Iss 1, Pp 1-8 (2021) |
| ISSN: | 2041-1723 |
| DOI: | 10.1038/s41467-021-26469-4 |
| Popis: | Class 2 CRISPR systems are exceptionally diverse, nevertheless, all share a single effector protein that contains a conserved RuvC-like nuclease domain. Interestingly, the size of these CRISPR-associated (Cas) nucleases ranges from >1000 amino acids (aa) for Cas9/Cas12a to as small as 400-600 aa for Cas12f. For in vivo genome editing applications, compact RNA-guided nucleases are desirable and would streamline cellular delivery approaches. Although miniature Cas12f effectors have been shown to cleave double-stranded DNA, targeted DNA modification in eukaryotic cells has yet to be demonstrated. Here, we biochemically characterize two miniature type V-F Cas nucleases, SpCas12f1 (497 aa) and AsCas12f1 (422 aa), and show that SpCas12f1 functions in both plant and human cells to produce targeted modifications with outcomes in plants being enhanced with short heat pulses. Our findings pave the way for the development of miniature Cas12f1-based genome editing tools. Miniature Cas12f editing systems are well suited for in vivo editing applications. Here the authors characterize the intrinsic activity of SpCas12f1 in plant and animal cells. |
| Databáze: | OpenAIRE |
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