journal article·2022-07Open access

A chemical CRISPR off switch efficiently controls gene editing

Xing-Yu Liu, Wei Xiong, Qian-Qian Qi, Hui-Min Ji, Yu-Tong Zhang, Hua-Jun Lei, Jian Liu, Ping Yin, Tian Tian, Xiang Zhou

Cell Reports Physical Science· Volume 3· Issue 7

Abstract

Chemical methods for regulating CRISPR activity hold great potential for spatiotemporal control of gene editing technologies. Here we report a small-molecule strategy to switch off CRISPR functionality. We develop clickable RNAs by modifying RNAs with an azido-containing functionality, which can be covalently conjugated in a second step with a complementary reaction partner, DBCO (dibenzocyclooctyne). We show that multimeric DBCO behaves quite differently than monoDBCO by imposing steric and topological constraints on RNAs. This generalizable strategy lays a solid foundation for developing clickable CRISPR off switches. We also show that this strategy works well for terminating CRISPR-Cas9-mediated genome editing in human cells.
Source: openalex_inverted_index · Quality: baseline

Research context

OpenAlex topics: CRISPR and Genetic Engineering · RNA and protein synthesis mechanisms · Advanced biosensing and bioanalysis techniques

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

References (45)

Show source-provided references
1. The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA10.1038/nature09523 ↗
2. CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III10.1038/nature09886 ↗
3. Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria10.1073/pnas.1208507109 ↗
4. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity10.1126/science.1225829 ↗
5. Multiplex genome engineering using CRISPR/Cas systems10.1126/science.1231143 ↗
6. RNA-guided human genome engineering via Cas910.1126/science.1232033 ↗
7. Naturally occurring off-switches for CRISPR-Cas910.1016/j.cell.2016.11.017 ↗
8. High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells10.1038/nbt.2623 ↗
9. Small-molecule control of protein function through Staudinger reduction10.1038/nchem.2573 ↗
10. Optical control of CRISPR/Cas9 gene editing10.1021/ja512664v ↗
11. Photoswitchable gRNAs for spatiotemporally controlled CRISPR-Cas-based genomic regulation10.1021/acscentsci.9b01093 ↗
12. Development of light-activated CRISPR using guide RNAs with photocleavable protectors10.1002/ange.201606123 ↗
13. Small molecule-triggered Cas9 protein with improved genome-editing specificity10.1038/nchembio.1793 ↗
14. Anti-CRISPR protein applications: natural brakes for CRISPR-Cas technologies10.1038/s41592-020-0771-6 ↗
15. Anti-CRISPRs: protein inhibitors of CRISPR-Cas systems10.1146/annurev-biochem-011420-111224 ↗
16. Potent CRISPR-Cas9 inhibitors from Staphylococcus genomes10.1073/pnas.1917668117 ↗
17. A high-throughput platform to identify small-molecule inhibitors of CRISPR-Cas910.1016/j.cell.2019.04.009 ↗
18. CRISPR-Switch regulates sgRNA activity by Cre recombination for sequential editing of two loci10.1038/s41467-019-13403-y ↗
19. A broadly applicable cross-linker for aliphatic polymers containing C-H bonds10.1126/science.aay6230 ↗
20. Click-ExM enables expansion microscopy for all biomolecules10.1038/s41592-020-01005-2 ↗
21. The chemistry and applications of RNA 2'-OH acylation10.1038/s41570-019-0147-6 ↗
22. RNA Cloaking by reversible acylation10.1002/ange.201708696 ↗
23. RNA control by photoreversible acylation10.1021/jacs.7b12408 ↗
24. Genetically encoded norbornene directs site-specific cellular protein labelling via a rapid bioorthogonal reaction10.1038/nchem.1250 ↗
25. Generation of a caged lentiviral vector through an unnatural amino acid for photo-switchable transduction10.1093/nar/gkz659 ↗
26. Strain-promoted "click" modification of a mesoporous metal-organic framework10.1021/ja307713q ↗
27. Strain-promoted copper-free "click" chemistry for 18F radiolabeling of bombesin10.1002/anie.201105547 ↗
28. Broccoli: rapid selection of an RNA mimic of green fluorescent protein by fluorescence-based selection and directed evolution10.1021/ja508478x ↗
29. Cas9 specifies functional viral targets during CRISPR-Cas adaptation10.1038/nature14245 ↗
30. SLX4IP antagonizes promiscuous BLM activity during ALT maintenance10.1016/j.molcel.2019.07.010 ↗
31. High-throughput screening of a CRISPR/Cas9 library for functional genomics in human cells10.1038/nature13166 ↗
32. CRISPR/Cas9-Mediated scanning for regulatory elements required for HPRT1 expression via thousands of large, programmed genomic deletions10.1016/j.ajhg.2017.06.010 ↗
33. Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection10.1038/nature19802 ↗
34. Nucleic acid detection with CRISPR-Cas13a/C2c210.1126/science.aam9321 ↗
35. Amplification-free detection of SARS-CoV-2 with CRISPR-Cas13a and mobile phone microscopy10.1016/j.cell.2020.12.001 ↗
36. The molecular architecture for RNA-guided RNA cleavage by Cas13a10.1016/j.cell.2017.06.050 ↗
37. A phage-encoded anti-CRISPR enables complete evasion of type VI-A CRISPR-Cas immunity10.1126/science.abb6151 ↗
38. Multiplexed CRISPR technologies for gene editing and transcriptional regulation10.1038/s41467-020-15053-x ↗
39. Electrophilic activity-based RNA probes reveal a self-alkylating RNA for RNA labeling10.1038/nchembio.1655 ↗
40. Programmable m(6)A modification of cellular RNAs with a Cas13-directed methyltransferase10.1038/s41587-020-0572-6 ↗
41. meCLICK-seq, a substrate-hijacking and RNA degradation strategy for the study of RNA methylation10.1021/acscentsci.0c01094 ↗
42. A walk along DNA using bipedal migration of a dynamic and covalent crosslinker10.1038/ncomms6591 ↗
43. Allosteric inhibition of CRISPR-Cas9 by bacteriophage-derived peptides10.1186/s13059-020-01956-x ↗
44. Coupling Cas9 to artificial inhibitory domains enhances CRISPR-Cas9 target specificity10.1126/sciadv.aay0187 ↗
45. Genome engineering using the CRISPR-Cas9 system10.1038/nprot.2013.143 ↗