ArticleNucleic acids research2020
Good guide, bad guide: spacer sequence-dependent cleavage efficiency of Cas12a.
Article in Nucleic acids research, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 60 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
60 citing papers in PubMed, 103 citations in OpenAlex.
- Development and clinical validation of a multiplex visual POCT platform based on RT-RAA-CRISPR/Cas12a for porcine diarrheal viruses.Synthetic and systems biotechnology · 2026Article
- Genetic transformation and CRISPR/Cas12a-mediated gene editing of European beech (Fagus sylvatica L.) employing a transient protoplast system.Communications biology · 2026Article
- Rapid detection ofMicrobiology spectrum · 2026Article
- DeepCas12a: a hybrid deep learning framework for accurate AsCas12a efficiency prediction from sequence and epigenetic information.BMC genomics · 2026Article
- Operating CRISPR/Cas12a in a complex nucleic acid sequence background.Nucleic acids research · 2026Article
- One-pot CRISPR-based point of care platform for rapid, specific and sensitive detection of HPV 16 without pre-amplification.Microsystems & nanoengineering · 2026Article
- Rapid and Sensitive Detection ofBiosensors · 2026Article
- Triplex DNA clamp regulates Cas12a activation for ssDNA and RNA sensing.Nucleic acids research · 2026Article
- Rapid and Sensitive Detection ofHuman mutation · 2026Article
- Article
- Advancements and Applications of Split Technology in CRISPR/Cas12a: Transforming Molecular Diagnostics and Biosensing.Biosensors · 2025Review
- Dynamic basis of supercoiling-dependent DNA interrogation by Cas12a via R-loop intermediates.Nature communications · 2025Article
- DNA target binding-induced pre-crRNA processing in type II and V CRISPR-Cas systems.Nucleic acids research · 2025Article
- 'Splice-at-will' Cas12a crRNA engineering enabled direct quantification of ultrashort RNAs.Nucleic acids research · 2025Article
- Disparate mechanisms counteract extraneous CRISPR RNA production in type II-C CRISPR-Cas systems.microLife · 2025Article
- Interpreting CRISPR-Cas12a enzyme kinetics through free energy change of nucleic acids.Nucleic acids research · 2024Article
- End-point diagnostics of Giardia duodenalis assemblages A and B by combining RPA with CRISPR/Cas12a from human fecal samples.Parasites & vectors · 2024Article
- The rise and future of CRISPR-based approaches for high-throughput genomics.FEMS microbiology reviews · 2024Review
- Kinetic dissection of pre-crRNA binding and processing by CRISPR-Cas12a.RNA (New York, N.Y.) · 2024Article
- Development of RPA-Cas12a assay for rapid and sensitive detection of Pneumocystis jirovecii.BMC microbiology · 2024Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
8 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Genome editing has recently made a revolutionary development with the introduction of the CRISPR-Cas technology. The programmable CRISPR-associated Cas9 and Cas12a nucleases generate specific dsDNA breaks in the genome, after which host DNA-repair mechanisms can be manipulated to implement the desired editing. Despite this spectacular progress, the efficiency of Cas9/Cas12a-based engineering can still be improved. Here, we address the variation in guide-dependent efficiency of Cas12a, and set out to reveal the molecular basis of this phenomenon. We established a sensitive and robust in vivo targeting assay based on loss of a target plasmid encoding the red fluorescent protein (mRFP). Our results suggest that folding of both the precursor guide (pre-crRNA) and the mature guide (crRNA) have a major influence on Cas12a activity. Especially, base pairing of the direct repeat, other than with itself, was found to be detrimental to the activity of Cas12a. Furthermore, we describe different approaches to minimize base-pairing interactions between the direct repeat and the variable part of the guide. We show that design of the 3' end of the guide, which is not involved in target strand base pairing, may result in substantial improvement of the guide's targeting potential and hence of its genome editing efficiency.
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Registered trials
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.