Diverse Mechanisms of CRISPR-Cas9 Inhibition by Type II Anti-CRISPR Proteins

Hwang, Sungwon; Maxwell, Karen L.

doi:10.1016/j.jmb.2023.168041

Cited by 13 publications

(6 citation statements)

References 72 publications

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“…The CRISPR/Cas technique is a new and powerful technology that, as a precise tool without the need for complex engineering, performs gene editing to selectively modify DNA sequences at any specific location in the genome with much greater precision than other genome editing technologies such as TALEN and ZFN [1][2][3] . With the discovery of this system, it has created a revolution in the development of cell-molecular biology along with genetics with profitable applications in biotechnology and medicine 4 .…”

Section: Introductionmentioning

confidence: 99%

CRISPR and Artificial intelligence to improve precision medicine: Future Perspectives and Potential Limitations

khoshandam¹,

Khoshandam

Soltaninejad

et al. 2023

Preprint

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The CRISPR/Cas9 genome editing system is a unique and new technology that allows genetics and medical researchers to modify or edit parts of the genome. This is done by deleting, inserting or changing parts of the DNA sequence. This method is currently the simplest, most widely used, and the most accurate method of genetic manipulation. This system has great potential to treat a wide range of genetic diseases. However, more research is needed to determine the advantages and disadvantages of the CRISPR system and establish best practices. On the other hand, individual patient treatment is one of the main goals of the medical field. Achieving this goal has been elusive due to a complex set of factors affecting disease and health. This work highlights key advances in the development of enabling technologies that advance the goal of personalized and precision medicine. Artificial intelligence is a simulation of human intelligence for computers, and is used in the interface of machines that is programmed to think and behave like a human. This review summarizes recent developments in the artificial intelligence (AI), CRISPR/Cas9, and examining clinical trials and potential based on these two in the advancement and development of precision and personalized medicine, cancer treatment and current and future challenges. Finally, the application of (AI) in the modification of the CRISPR/Cas9 system is highlighted.

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Section: Introductionmentioning

confidence: 99%

CRISPR and Artificial intelligence to improve precision medicine: Future Perspectives and Potential Limitations

khoshandam¹,

Khoshandam

Soltaninejad

et al. 2023

Preprint

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“…Moreover, both AcrIIC1 and AcrIIA5 are broad-spectrum Acrs, which can inhibit a wide range of Cas9 orthologs by preventing target DNA cleavage ( 26–28 ). AcrIIC1 binds to the HNH endonuclease domain of Cas9 to prevent conformational changes for DNA cleavage within Cas9, while the precise mechanism of AcrIIA5 remains unresolved ( 19 , 28 ). Additionally, only a small portion of anti-CRISPR proteins are developed for genome editing and modulation, including AcrIIA2, AcrIIA4, AcrIIC1, AcrIIC3, AcrIIA25.1 and AcrIIA32.1 ( 19 , 20 ).…”

Section: Introductionmentioning

confidence: 99%

“…To date, elucidated Acr proteins mainly affect the binding of CRISPR-Cas surveillance complex to target DNA (or RNA), like AcrIIA4, AcrIIA27 and AcrIIA31, etc ( 19 , 28 ). CRISPR interference (CRISPRi) assays for gene repression by Cas effectors have been adopted to investigate whether Acrs can affect DNA recognition of CRISPR-Cas systems, which is an effective method to reveal the inhibitory strategies of Acrs ( 25 , 27 , 37 , 38 ).…”

Section: Introductionmentioning

confidence: 99%

Rapid characterization of anti-CRISPR proteins and optogenetically engineered variants using a versatile plasmid interference system

Song,

Tian,

et al. 2023

Nucleic Acids Research

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Anti-CRISPR (Acr) proteins are encoded by mobile genetic elements to overcome the CRISPR immunity of prokaryotes, displaying promises as controllable tools for modulating CRISPR-based applications. However, characterizing novel anti-CRISPR proteins and exploiting Acr-related technologies is a rather long and tedious process. Here, we established a versatile plasmid interference with CRISPR interference (PICI) system in Escherichia coli for rapidly characterizing Acrs and developing Acr-based technologies. Utilizing the PICI system, we discovered two novel type II-A Acrs (AcrIIA33 and AcrIIA34), which can inhibit the activity of SpyCas9 by affecting DNA recognition of Cas9. We further constructed a circularly permuted AcrIIA4 (cpA4) protein and developed optogenetically engineered, robust AcrIIA4 (OPERA4) variants by combining cpA4 with the light-oxygen-voltage 2 (LOV2) blue light sensory domain. OPERA4 variants are robust light-dependent tools for controlling the activity of SpyCas9 by approximately 1000-fold change under switching dark-light conditions in prokaryotes. OPERA4 variants can achieve potent light-controllable genome editing in human cells as well. Together, our work provides a versatile screening system for characterizing Acrs and developing the Acr-based controllable tools.

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“…The programmable nature of Cas effector proteins (including Cas9 and Cas12a) has facilitated their utilization for applications ranging from nucleic acid detection to genome editing ( 4 , 5 ). Viruses encode anti-CRISPR (Acr) proteins to evade CRISPR-Cas immunity ( 6 – 8 ). Acr proteins can have different evolutionary origins and allow escape from CRISPR-Cas immunity through distinct mechanisms.…”

mentioning

confidence: 99%

“…Cas9-mediated DNA cleavage can be inhibited by Acr proteins by preventing crRNA loading, blocking DNA binding, or inhibiting DNA cleavage ( 8 ). To date, six different type II-C-specific inhibitors have been identified (AcrIIC1-AcrIIC6): AcrIIC1 and AcrIIC3 inhibit the nuclease activity of HNH, AcrIIC2 prevents crRNA loading, and AcrIIC5 and AcrIIC6 inhibit DNA binding ( 8 ).…”

mentioning

confidence: 99%

Keeping Cas9 out of the (R-)loop

Swarts

2023

Proc. Natl. Acad. Sci. U.S.A.

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Diverse Mechanisms of CRISPR-Cas9 Inhibition by Type II Anti-CRISPR Proteins

Cited by 13 publications

References 72 publications

CRISPR and Artificial intelligence to improve precision medicine: Future Perspectives and Potential Limitations

CRISPR and Artificial intelligence to improve precision medicine: Future Perspectives and Potential Limitations

Rapid characterization of anti-CRISPR proteins and optogenetically engineered variants using a versatile plasmid interference system

Keeping Cas9 out of the (R-)loop

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