A chemical CRISPR off switch efficiently controls gene editing

Liu, Xingyu; Xiong, Wei; Qi, Qian-Qian; Ji, Huimin; Zhang, Yutong; Lei, Huajun; Liu, Jian; Yin, Ping; Tian, Tian; Zhou, Xiang

doi:10.1016/j.xcrp.2022.100956

Cited by 5 publications

(5 citation statements)

References 45 publications

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“…After 24 h, the efficiency of the gene editing (insertion/deletion (indel) mutation) was evaluated by a T7EI nuclease assay. [ 23 ] Before performing conditional control gene editing in live cells, we first assessed the cytotoxicity of DPPEA to Hela‐OC cells. We observed that DPPEA caused no toxicity in cells up to 512 µ m (Figure S16 , Supporting Information).…”

Section: Resultsmentioning

confidence: 99%

Chemical Control of CRISPR Gene Editing via Conditional Diacylation Crosslinking of Guide RNAs

Lei

Zeng

et al. 2023

Advanced Science

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Conditional control of RNA structure and function has emerged as an effective toolkit. Here, a strategy based on a one‐step introduction of diacylation linkers and azide groups on the 2′‐OH of RNA is advance. Selected from eight phosphine reagents, it is found that 2‐(diphenylphosphino)ethylamine has excellent performance in reducing azides via a Staudinger reduction to obtain the original RNA. It is demonstrated that the enzymatic activities of Cas13 and Cas9 can be regulated by chemically modified guide RNAs, and further achieved ligand‐induced gene editing in living cells by a controllable CRISPR/Cas9 system.

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Section: Resultsmentioning

confidence: 99%

Chemical Control of CRISPR Gene Editing via Conditional Diacylation Crosslinking of Guide RNAs

Lei

Zeng

et al. 2023

Advanced Science

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“…Such modified CRISPR/Cas systems can be activated or deactivated through controlled operations, thereby averting any potential toxic side effects stemming from excessive gene editing. Our previous work has explored anti‐CRISPR strategies grounded in supramolecular chemistry, [ 18 ] click chemistry, [ 19 ] and gRNA modification. [ 20‐21 ] However, the continued development of novel Cas9 regulatory strategies with diverse mechanisms will expand the potential applications of Cas9 in the future.…”

Section: Background and Originality Contentmentioning

confidence: 99%

Regulating CRISPR/Cas9 Using Streptavidin‐Biotin Interactions^†

Shen,

Xiong,

et al. 2024

Chin. J. Chem.

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Comprehensive SummaryCurrently, CRISPR/Cas9 technology has found widespread applications across various domains. However, the utility of CRISPR/Cas9 is encumbered by issues pertaining to its reliability and safety, primarily stemming from the uncontrolled activity of the system. Therefore, the design and development of CRISPR/Cas9 systems with controllable activity is of paramount importance. Biotin, characterized by its small molecular weight, and streptavidin, distinguished by its substantial spatial steric hindrance, can be harnessed as an ideal OFF switch (termed a "bioactivity brake") due to their interaction characteristics. In this work, we present a strategy that employs the streptavidin‐biotin interaction as a "brake system" for CRISPR/Cas9, effectively allowing for the shutdown of the enzymatic activity of CRISPR/Cas9.

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“…The same research group reported clickable gRNAs that are chemically labeled to contain azido groups [ 94 ]. The labeled gRNAs are still active due to the small-sized chemical modification.…”

Section: Small-molecule Control Of Grnasmentioning

confidence: 99%

“…Molecules having two or three DBCO groups induced crosslinking between gRNAs and displayed higher potency than those having a single DBCO. This strategy was also demonstrated in human cells to inhibit genome editing, and it was expandable to the crRNA of Cas13a ( Figure 5 E) [ 94 ].…”

Section: Small-molecule Control Of Grnasmentioning

confidence: 99%

Small Molecules for Enhancing the Precision and Safety of Genome Editing

2022

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Clustered regularly interspaced short palindromic repeats (CRISPR)-based genome-editing technologies have revolutionized biology, biotechnology, and medicine, and have spurred the development of new therapeutic modalities. However, there remain several barriers to the safe use of CRISPR technologies, such as unintended off-target DNA cleavages. Small molecules are important resources to solve these problems, given their facile delivery and fast action to enable temporal control of the CRISPR systems. Here, we provide a comprehensive overview of small molecules that can precisely modulate CRISPR-associated (Cas) nucleases and guide RNAs (gRNAs). We also discuss the small-molecule control of emerging genome editors (e.g., base editors) and anti-CRISPR proteins. These molecules could be used for the precise investigation of biological systems and the development of safer therapeutic modalities.

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A chemical CRISPR off switch efficiently controls gene editing

Cited by 5 publications

References 45 publications

Chemical Control of CRISPR Gene Editing via Conditional Diacylation Crosslinking of Guide RNAs

Chemical Control of CRISPR Gene Editing via Conditional Diacylation Crosslinking of Guide RNAs

Regulating CRISPR/Cas9 Using Streptavidin‐Biotin Interactions^†

Small Molecules for Enhancing the Precision and Safety of Genome Editing

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A chemical CRISPR off switch efficiently controls gene editing

Cited by 5 publications

References 45 publications

Chemical Control of CRISPR Gene Editing via Conditional Diacylation Crosslinking of Guide RNAs

Chemical Control of CRISPR Gene Editing via Conditional Diacylation Crosslinking of Guide RNAs

Regulating CRISPR/Cas9 Using Streptavidin‐Biotin Interactions†

Small Molecules for Enhancing the Precision and Safety of Genome Editing

Contact Info

Product

Resources

About

Regulating CRISPR/Cas9 Using Streptavidin‐Biotin Interactions^†