ArticleNucleic acids research2022
Efficient target cleavage by Type V Cas12a effectors programmed with split CRISPR RNA.
Article in Nucleic acids research, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
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Who cites it
35 citing papers in PubMed, 75 citations in OpenAlex.
- Molecular mechanisms and biotechnology applications of CRISPR-Cas12a.Nature reviews. Molecular cell biology · 2026Review
- Molecular engineering of CRISPR/Cas12a: from activity enhancement to exponential signal amplification.Chemical science · 2026Review
- DNA-guided CRISPR-Cas12 for cellular RNA targeting.Nature biotechnology · 2026Article
- crRNA Truncation MinimizesJACS Au · 2026Article
- Activation mechanism and integrated one-pot assay of chiral-like cRNA-enhanced CRISPR/Cas12a systems.Nature communications · 2026Article
- Advanced CRISPR Technologies for RNA Imaging in Live Cells.Chemical & biomedical imaging · 2026Review
- A novel Dual-guide CRISPR-Cas13 strategy improves specificity for single-nucleotide variant detection.Nucleic acids research · 2026Article
- crRNA scaffold remodeling controls CRISPR-Cas12a activity for enhanced performance.Nucleic acids research · 2026Article
- Programmable hooded DNA switches for conditional control of CRISPR/Cas12a in multiplexed biosensing.Journal of nanobiotechnology · 2026Article
- Article
- Cas12a-assisted split crRNA complex for analysis and detection of diverse entities.Nucleic acids research · 2025Article
- Advancements and Applications of Split Technology in CRISPR/Cas12a: Transforming Molecular Diagnostics and Biosensing.Biosensors · 2025Review
- AsCas12a tolerates insertions in target DNA.Nucleic acids research · 2025Article
- Identification of a key nucleotide influencing Cas12a crRNA activity for universal photo-controlled CRISPR diagnostics.Nature communications · 2025Article
- Regulating cleavage activity and enabling microRNA detection with split sgRNA in Cas12b.Nature communications · 2025Article
- Enhancement of CRISPR-Cas12a system through universal circular RNA design.Cell reports methods · 2025Article
- Spatially blocked split CRISPR-Cas12a system for ultra-sensitive and versatile small molecule activation and detection.Nature communications · 2025Article
- Nonequilibrium hybridization-driven CRISPR/Cas adapter with extended energetic penalty for discrimination of single-nucleotide variants.Nucleic acids research · 2025Article
- Targeted deletions in human mitochondrial DNA engineered by Type V CRISPR-Cas12a system.NAR molecular medicine · 2025Article
- Expanding Cas12a Activity Control with an RNA G-Quadruplex at the 5' end of CRISPR RNA.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
13 authors at 5 institutions in 4 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
CRISPR RNAs (crRNAs) that direct target DNA cleavage by Type V Cas12a nucleases consist of constant repeat-derived 5'-scaffold moiety and variable 3'-spacer moieties. Here, we demonstrate that removal of most of the 20-nucleotide scaffold has only a slight effect on in vitro target DNA cleavage by a Cas12a ortholog from Acidaminococcus sp. (AsCas12a). In fact, residual cleavage was observed even in the presence of a 20-nucleotide crRNA spacer moiety only. crRNAs split into separate scaffold and spacer RNAs catalyzed highly specific and efficient cleavage of target DNA by AsCas12a in vitro and in lysates of human cells. In addition to dsDNA target cleavage, AsCas12a programmed with split crRNAs also catalyzed specific ssDNA target cleavage and non-specific ssDNA degradation (collateral activity). V-A effector nucleases from Francisella novicida (FnCas12a) and Lachnospiraceae bacterium (LbCas12a) were also functional with split crRNAs. Thus, the ability of V-A effectors to use split crRNAs appears to be a general property. Though higher concentrations of split crRNA components are needed to achieve efficient target cleavage, split crRNAs open new lines of inquiry into the mechanisms of target recognition and cleavage and may stimulate further development of single-tube multiplex and/or parallel diagnostic tests based on Cas12a nucleases.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.