ArticleNature communications2021
Gene editing with CRISPR-Cas12a guides possessing ribose-modified pseudoknot handles.
Article in Nature communications, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- Molecular mechanisms and biotechnology applications of CRISPR-Cas12a.Nature reviews. Molecular cell biology · 2026Review
- Single-residue engineering of Klenow fragment enables broad acceptance of chemically modified nucleotides.Nucleic acids research · 2026Article
- A nanopore-based HIV-1 reference epitranscriptome.Nucleic acids research · 2026Article
- Chemical control of 2'-hydroxyl-dependent Cas9 target engagement enables CRISPR RNA ribose replacement.bioRxiv : the preprint server for biology · 2026Article
- Effects of NBioconjugate chemistry · 2025Article
- LbuCas13a directly targets DNA and elicits strong trans-cleavage activity.Nature biomedical engineering · 2025Article
- RNA chemistry and therapeutics.Nature reviews. Drug discovery · 2025Review
- A solid-phase enzymatic synthesis platform for the facile production of 2'-fluoroarabinonucleic acid (FANA) and chimeric XNA oligonucleotides using an evolved XNA polymerase.Nucleic acids research · 2025Article
- Chemical engineering of CRISPR-Cas systems for therapeutic application.Nature reviews. Drug discovery · 2025Review
- Engineering a DNA polymerase for modifying large RNA at specific positions.Nature chemistry · 2025Article
- Recent advances in nanoparticulate RNA delivery systems.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
- Asymmetric CRISPR enabling cascade signal amplification for nucleic acid detection by competitive crRNA.Nature communications · 2023Article
- Regulation of the CRISPR-Cas12a system by methylation and demethylation of guide RNA.Chemical science · 2023Article
- Chemical Modifications of CRISPR RNAs to Improve Gene-Editing Activity and Specificity.Journal of the American Chemical Society · 2022Review
Corrections and comments
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Authors and funding
8 authors.
Funding
Abstract
CRISPR-Cas12a is a leading technology for development of model organisms, therapeutics, and diagnostics. These applications could benefit from chemical modifications that stabilize or tune enzyme properties. Here we chemically modify ribonucleotides of the AsCas12a CRISPR RNA 5' handle, a pseudoknot structure that mediates binding to Cas12a. Gene editing in human cells required retention of several native RNA residues corresponding to predicted 2'-hydroxyl contacts. Replacing these RNA residues with a variety of ribose-modified nucleotides revealed 2'-hydroxyl sensitivity. Modified 5' pseudoknots with as little as six out of nineteen RNA residues, with phosphorothioate linkages at remaining RNA positions, yielded heavily modified pseudoknots with robust cell-based editing. High trans activity was usually preserved with cis activity. We show that the 5' pseudoknot can tolerate near complete modification when design is guided by structural and chemical compatibility. Rules for modification of the 5' pseudoknot should accelerate therapeutic development and be valuable for CRISPR-Cas12a diagnostics.
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