ArticleCell regeneration (London, England)2019
Efficient cleavage resolves PAM preferences of CRISPR-Cas in human cells.
Article in Cell regeneration (London, England), 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 37 citations in OpenAlex.
- Nature Inspired Delivery Vehicles for CRISPR-Based Genome Editing.Small (Weinheim an der Bergstrasse, Germany) · 2026Review
- Uncovering Cas9 PAM diversity through metagenomic mining and machine learning.Nature communications · 2026Article
- GenomePAM directs PAM characterization and engineering of CRISPR-Cas nucleases using mammalian genome repeats.Nature biomedical engineering · 2026Article
- Precision gene editing: The power of CRISPR-Cas in modern genetics.Molecular therapy. Nucleic acids · 2025Review
- PAM-readID is a rapid, simple, and accurate PAM determination method for CRISPR-Cas enzymes in mammalian cells.Communications biology · 2025Article
- GenomePAM directs PAM characterization and engineering of CRISPR-Cas nucleases using mammalian genome repeats.Research square · 2025Article
- Engineering miniature IscB nickase for robust base editing with broad targeting range.Nature chemical biology · 2024Article
- Emerging CRISPR Therapies for Precision Gene Editing and Modulation in the Cardiovascular Clinic.Current cardiology reports · 2024Review
- Protocol for Allele-Specific Epigenome Editing Using CRISPR/dCas9.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Development of super-specific epigenome editing by targeted allele-specific DNA methylation.Epigenetics & chromatin · 2023Article
- Hybrid Multitask Learning Reveals Sequence Features Driving Specificity in the CRISPR/Cas9 System.Biomolecules · 2023Article
- Microbiome engineering for bioremediation of emerging pollutants.Bioprocess and biosystems engineering · 2023Review
- Versatile and efficient genome editing with Neisseria cinerea Cas9.Communications biology · 2022Article
- Synergistic engineering of CRISPR-Cas nucleases enables robust mammalian genome editing.Innovation (Cambridge (Mass.)) · 2022Article
- Can SpRY recognize any PAM in human cells?Journal of Zhejiang University. Science. B · 2022Article
- Versatile and efficient in vivo genome editing with compact Streptococcus pasteurianus Cas9.Molecular therapy : the journal of the American Society of Gene Therapy · 2022Article
- Combination of Isothermal Recombinase-Aided Amplification and CRISPR-Cas12a-Mediated Assay for Rapid Detection of Major Severe Acute Respiratory Syndrome Coronavirus 2 Variants of Concern.Frontiers in microbiology · 2022Article
- Activation of HERV-K(HML-2) disrupts cortical patterning and neuronal differentiation by increasing NTRK3.Cell stem cell · 2021Article
- Article
- Welcome to Cell Regeneration.Cell regeneration (London, England) · 2020Article
Corrections and comments
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Authors and funding
13 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Clustered regularly interspaced short palindromic repeats and associated proteins (CRISPR-Cas) of bacterial adaptive immunity have been adopted as a powerful and versatile tool for manipulation of the genome. This paradigm has been widely applied in biological research and treatments of animal or cellular disease models. A critical feature of CRISPR-Cas is the protospacer adjacent motif (PAM), which dictates the DNA target recognition mechanism of Cas proteins. While, direct identifying functional PAM sequences in human cells remains a challenge. Here, we developed a positive screen system termed PAM-DOSE (PAM Definition by Observable Sequence Excision) to delineate the functional PAMs in human cells. Specifically, the PAM libraries for CRISPR-Cas (SpCas9, SpCas9-NG, FnCas12a, AsCas12a, LbCas12a and MbCas12a) were generated and the corresponding CRISPR-Cas mediated cleaved fragments with functional PAM in human cells were harvested for DNA sequencing, which could be tracked and visualized with either florescence microscopy or flow cytometry analysis. With this system, we identified the functional PAMs of CRISPR-Cas members. We also found that spacer sequence affects the PAM preference of Cas proteins. This method will facilitate identification of functional PAMs for Cas-mediated human genome editing applications.
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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.