ArticleMolecular cell2022
Chimeric CRISPR-CasX enzymes and guide RNAs for improved genome editing activity.
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Article in Molecular cell, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 36 papers.
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36citing papers in PubMed
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Who cites it
36 citing papers in PubMed.
- Miniaturized CRISPR: Ultra Compact Systems for In Vivo Delivery and Portable Diagnostics.Annals of biomedical engineering · 2026Review
- Delivering the future of immunotherapy: A state-of-the-art review of gene editing in immune cells with lipid nanoparticles.Materials today. Bio · 2026Review
- Bridge helix of Cas12a is an allosteric regulator of R-loop formation and RuvC activation.Nature communications · 2026Article
- Comprehensive assessment of activity, specificity, and safety of hypercompact TnpB systems for gene editing.Genome biology · 2026Article
- Engineering TnpB as a compact RNA-guided genome editor from molecular constraints to design principles.Frontiers in genome editing · 2026Review
- Phage-associated Cas12p nucleases require binding to bacterial thioredoxin for activation and cleavage of target DNA.Nature microbiology · 2026Article
- Enhanced cleavage of genomicRNA biology · 2025Article
- Engineering a CRISPR-associated IscB system for developing miniature genome-editing tools in human cells and mouse embryos.Nature communications · 2025Article
- Dystrophin Restorative and Compensatory Gene Addition Therapies for Duchenne Muscular Dystrophy: Could CRISPRa Provide a Realistic Alternative?Muscles (Basel, Switzerland) · 2025Review
- Improving Gene Knock-In Efficiencies in Sheep Primary Cells Using a CRISPR/Cas9-Gal4 System.Current issues in molecular biology · 2025Article
- CRISPR-driven diagnostics: Molecular mechanisms, clinical efficacy and translational challenges.Clinical and translational medicine · 2025Review
- Recent advances in therapeutic gene-editing technologies.Molecular therapy : the journal of the American Society of Gene Therapy · 2025Review
- Nano-Polymers as Cas9 Inhibitors.Polymers · 2025Article
- Cas12e orthologs evolve variable structural elements to facilitate dsDNA cleavage.Nature communications · 2024Article
- CRISPR-Cas9-mediated homology-directed repair for precise gene editing.Molecular therapy. Nucleic acids · 2024Review
- Research Progress and Application of Miniature CRISPR-Cas12 System in Gene Editing.International journal of molecular sciences · 2024Review
- PlmCas12e Utilizes Glu662 to Prevent Cleavage Site Occupation by Positively Charged Residues Before Target Strand Cleavage.Molecules (Basel, Switzerland) · 2024Article
- Conformational dynamics of CasX (Cas12e) in mediating DNA cleavage revealed by single-molecule FRET.Nucleic acids research · 2024Article
- CRISPR technology in human diseases.MedComm · 2024Review
- Cas12a domain flexibility guides R-loop formation and forces RuvC resetting.Molecular cell · 2024Article
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Authors and funding
18 authors.
Connor A TsuchidaUniversity of California, Berkeley-University of California, San Francisco Graduate Program in Bioengineering, University of California, Berkeley, California 94720, USA; Innovative Genomics Institute, University of California, Berkeley, California 94720, USA.
Shouyue ZhangBeijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China.
Mohammad Saffari DoostInnovative Genomics Institute, University of California, Berkeley, California 94720, USA; California Institute for Quantitative Biosciences, University of California, Berkeley, California 94720, USA.
Yuqian ZhaoBeijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China.
Jia WangBeijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China.
Elizabeth O'BrienInnovative Genomics Institute, University of California, Berkeley, California 94720, USA; California Institute for Quantitative Biosciences, University of California, Berkeley, California 94720, USA.
Huan FangInstitute of Molecular Medicine, College of Future Technology, Peking University, Beijing 100871, China.
Cheng-Ping LiBeijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China.
Danyuan LiBeijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China.
Zhuo-Yan HaiBeijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China.
Jonathan ChuckDepartment of Integrative Structural and Computational Biology, The Scripps Research Institute, Jupiter, Florida 33458, USA.
Julian BrötzmannDepartment of Structural Cell Biology, Max Planck Institute of Biochemistry, Martinsried, Germany.
Araz VartoumianInnovative Genomics Institute, University of California, Berkeley, California 94720, USA; California Institute for Quantitative Biosciences, University of California, Berkeley, California 94720, USA.
David BursteinSchool of Molecular Cell Biology and Biotechnology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 6997801, Israel.
Xiao-Wei ChenInstitute of Molecular Medicine, College of Future Technology, Peking University, Beijing 100871, China.
Eva NogalesCalifornia Institute for Quantitative Biosciences, University of California, Berkeley, California 94720, USA; Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA; Molecular Biophysics & Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA; Howard Hughes Medical Institute, University of California, Berkeley, California 94720, USA.
Jennifer A DoudnaUniversity of California, Berkeley-University of California, San Francisco Graduate Program in Bioengineering, University of California, Berkeley, California 94720, USA; Innovative Genomics Institute, University of California, Berkeley, California 94720, USA; California Institute for Quantitative Biosciences, University of California, Berkeley, California 94720, USA; Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA; Molecular Biophysics & Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA; Howard Hughes Medical Institute, University of California, Berkeley, California 94720, USA; Department of Chemistry, University of California, Berkeley, California 94720, USA; Gladstone Institute of Data Science and Biotechnology. Gladstone Institutes, San Francisco, California 94158, USA. Electronic address: doudna@berkeley.edu.
Jun-Jie Gogo LiuBeijing Advanced Innovation Center for Structural Biology & Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, Beijing 100084, China. Electronic address: junjiegogoliu@tsinghua.edu.cn.
Funding
TUBULIN STRUCTURE BY ELECTRON CRYSTALLOGRAPHYP01GM051487 · NIGMS · UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB · PI NOGALES, EVA · 1994 to 2018
$27.6MCharacterization and engineering of a compact genome editor for adeno-associated virus delivery to correct mutations causing cystic fibrosisF31HL156468 · NHLBI · UNIVERSITY OF CALIFORNIA BERKELEY · PI TSUCHIDA, CONNOR ANDREW · 2021 to 2022
$77kHoward Hughes Medical InstituteNHLBI NIH HHS F31 HL156468NIGMS NIH HHS P01 GM051487
Abstract
A compact protein with a size of <1,000 amino acids, the CRISPR-associated protein CasX is a fundamentally distinct RNA-guided nuclease when compared to Cas9 and Cas12a. Although it can induce RNA-guided genome editing in mammalian cells, the activity of CasX is less robust than that of the widely used S. pyogenes Cas9. Here, we show that structural features of two CasX homologs and their guide RNAs affect the R-loop complex assembly and DNA cleavage activity. Cryo-EM-based structural engineering of either the CasX protein or the guide RNA produced two new CasX genome editors (DpbCasX-R3-v2 and PlmCasX-R1-v2) with significantly improved DNA manipulation efficacy. These results advance both the mechanistic understanding of CasX and its application as a genome-editing tool.
Indexed as
Gene EditingRNA, Guide, CRISPR-Cas SystemsAnimalsCRISPR-Cas SystemsEndonucleasesMammalsRNAEndonucleasesRNARNA, Guide, CRISPR-Cas SystemsCas12eCasXCRISPRcryo-EMDNA cleavagegenome editingnucleic acid manipulationRNA-guided DNA nucleasesgRNAstructural engineering
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