Dynamic observations of CRISPR-Cas target recognition and cleavage heterogeneities
CRISPR-Cas systems (clustered regularly interspaced short palindromic repeats) have shown great potential as efficient gene editing tools in disease therapeutics. Although numerous CRISPR-Cas systems have been developed, detailed mechanisms of target recognition and DNA cleavage are still unclear. I...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
De Gruyter
2022-08-01
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| Series: | Nanophotonics |
| Subjects: | |
| Online Access: | https://doi.org/10.1515/nanoph-2022-0286 |
| _version_ | 1797947885696843776 |
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| author | Zhang Zhijia Jeong Haechan Zu Di Zhao Xintao Senaratne Pramith Filbin John Silber Brett Kang Sarah Gladstone Ann Lau Matthew Cui Guangjie Park Younggeun Lee Somin Eunice |
| author_facet | Zhang Zhijia Jeong Haechan Zu Di Zhao Xintao Senaratne Pramith Filbin John Silber Brett Kang Sarah Gladstone Ann Lau Matthew Cui Guangjie Park Younggeun Lee Somin Eunice |
| author_sort | Zhang Zhijia |
| collection | DOAJ |
| description | CRISPR-Cas systems (clustered regularly interspaced short palindromic repeats) have shown great potential as efficient gene editing tools in disease therapeutics. Although numerous CRISPR-Cas systems have been developed, detailed mechanisms of target recognition and DNA cleavage are still unclear. In this work, we dynamically observe the entire process of conjugation, target recognition and DNA cleavage by single particle spectroscopy of CRISPR-Cas systems on single particle surfaces (gold) with the unique advantage of extended time periods. We show the CRISPR-Cas system, comprised of Cas endonuclease and single guide RNA, is stable and functional on single particle surfaces. Owing to the photostability of single particle surfaces, we directly observe in real time the entire dynamic process of conjugation, target recognition and DNA cleavage without photobleaching. We find heterogeneity in target recognition and DNA cleavage processes in which individual spectra vary significantly from one another as well as from the ensemble. We believe an in depth understanding of heterogeneities in CRISPR-Cas systems can overcome potential barriers in precision medicine and personalized disease therapeutics. |
| first_indexed | 2024-04-10T21:34:46Z |
| format | Article |
| id | doaj.art-561ac2c02a8c4721af37f7a9d353f425 |
| institution | Directory Open Access Journal |
| issn | 2192-8614 |
| language | English |
| last_indexed | 2024-04-10T21:34:46Z |
| publishDate | 2022-08-01 |
| publisher | De Gruyter |
| record_format | Article |
| series | Nanophotonics |
| spelling | doaj.art-561ac2c02a8c4721af37f7a9d353f4252023-01-19T12:47:01ZengDe GruyterNanophotonics2192-86142022-08-0111194419442510.1515/nanoph-2022-0286Dynamic observations of CRISPR-Cas target recognition and cleavage heterogeneitiesZhang Zhijia0Jeong Haechan1Zu Di2Zhao Xintao3Senaratne Pramith4Filbin John5Silber Brett6Kang Sarah7Gladstone Ann8Lau Matthew9Cui Guangjie10Park Younggeun11Lee Somin Eunice12Department of Electrical & Computer Engineering, Biomedical Engineering, Applied Physics, Biointerfaces Institute, Macromolecular Science & Engineering, University of Michigan, Ann Arbor, USADepartment of Electrical & Computer Engineering, Biomedical Engineering, Applied Physics, Biointerfaces Institute, Macromolecular Science & Engineering, University of Michigan, Ann Arbor, USADepartment of Electrical & Computer Engineering, Biomedical Engineering, Applied Physics, Biointerfaces Institute, Macromolecular Science & Engineering, University of Michigan, Ann Arbor, USADepartment of Electrical & Computer Engineering, Biomedical Engineering, Applied Physics, Biointerfaces Institute, Macromolecular Science & Engineering, University of Michigan, Ann Arbor, USADepartment of Electrical & Computer Engineering, Biomedical Engineering, Applied Physics, Biointerfaces Institute, Macromolecular Science & Engineering, University of Michigan, Ann Arbor, USADepartment of Electrical & Computer Engineering, Biomedical Engineering, Applied Physics, Biointerfaces Institute, Macromolecular Science & Engineering, University of Michigan, Ann Arbor, USADepartment of Electrical & Computer Engineering, Biomedical Engineering, Applied Physics, Biointerfaces Institute, Macromolecular Science & Engineering, University of Michigan, Ann Arbor, USADepartment of Electrical & Computer Engineering, Biomedical Engineering, Applied Physics, Biointerfaces Institute, Macromolecular Science & Engineering, University of Michigan, Ann Arbor, USADepartment of Electrical & Computer Engineering, Biomedical Engineering, Applied Physics, Biointerfaces Institute, Macromolecular Science & Engineering, University of Michigan, Ann Arbor, USADepartment of Electrical & Computer Engineering, Biomedical Engineering, Applied Physics, Biointerfaces Institute, Macromolecular Science & Engineering, University of Michigan, Ann Arbor, USADepartment of Electrical & Computer Engineering, Biomedical Engineering, Applied Physics, Biointerfaces Institute, Macromolecular Science & Engineering, University of Michigan, Ann Arbor, USADepartment of Mechanical Engineering, University of Michigan, Ann Arbor, USADepartment of Electrical & Computer Engineering, Biomedical Engineering, Applied Physics, Biointerfaces Institute, Macromolecular Science & Engineering, University of Michigan, Ann Arbor, USACRISPR-Cas systems (clustered regularly interspaced short palindromic repeats) have shown great potential as efficient gene editing tools in disease therapeutics. Although numerous CRISPR-Cas systems have been developed, detailed mechanisms of target recognition and DNA cleavage are still unclear. In this work, we dynamically observe the entire process of conjugation, target recognition and DNA cleavage by single particle spectroscopy of CRISPR-Cas systems on single particle surfaces (gold) with the unique advantage of extended time periods. We show the CRISPR-Cas system, comprised of Cas endonuclease and single guide RNA, is stable and functional on single particle surfaces. Owing to the photostability of single particle surfaces, we directly observe in real time the entire dynamic process of conjugation, target recognition and DNA cleavage without photobleaching. We find heterogeneity in target recognition and DNA cleavage processes in which individual spectra vary significantly from one another as well as from the ensemble. We believe an in depth understanding of heterogeneities in CRISPR-Cas systems can overcome potential barriers in precision medicine and personalized disease therapeutics.https://doi.org/10.1515/nanoph-2022-0286bioplasmonicscasclustered regularly interspaced short palindromic repeatsgene editinggold nanorodplasmonicssgrna |
| spellingShingle | Zhang Zhijia Jeong Haechan Zu Di Zhao Xintao Senaratne Pramith Filbin John Silber Brett Kang Sarah Gladstone Ann Lau Matthew Cui Guangjie Park Younggeun Lee Somin Eunice Dynamic observations of CRISPR-Cas target recognition and cleavage heterogeneities Nanophotonics bioplasmonics cas clustered regularly interspaced short palindromic repeats gene editing gold nanorod plasmonics sgrna |
| title | Dynamic observations of CRISPR-Cas target recognition and cleavage heterogeneities |
| title_full | Dynamic observations of CRISPR-Cas target recognition and cleavage heterogeneities |
| title_fullStr | Dynamic observations of CRISPR-Cas target recognition and cleavage heterogeneities |
| title_full_unstemmed | Dynamic observations of CRISPR-Cas target recognition and cleavage heterogeneities |
| title_short | Dynamic observations of CRISPR-Cas target recognition and cleavage heterogeneities |
| title_sort | dynamic observations of crispr cas target recognition and cleavage heterogeneities |
| topic | bioplasmonics cas clustered regularly interspaced short palindromic repeats gene editing gold nanorod plasmonics sgrna |
| url | https://doi.org/10.1515/nanoph-2022-0286 |
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