ArticleScientific reports2019
A qPCR method for genome editing efficiency determination and single-cell clone screening in human cells.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed, 33 citations in OpenAlex.
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- Engineering with EP2/EP4 knockout and IL-15 transpresentation renders stem cell-derived NK cells self-persistent and resistant to PGE2 inhibition.Frontiers in immunology · 2026Article
- DR1 activates histone gene expression to maintain pancreatic cancer cell survival through the ATAC complex.Cancer gene therapy · 2025Article
- Comprehensive benchmarking of genome editing quantification methods for plant applications.iScience · 2025Article
- Lentiviral Transduction-based CRISPR/Cas9 Editing ofCurrent genomics · 2023Article
- A Digital PCR Method Based on Highly Specific Taq for Detecting Gene Editing and Mutations.International journal of molecular sciences · 2023Article
- Loss of RREB1 in pancreatic beta cells reduces cellular insulin content and affects endocrine cell gene expression.Diabetologia · 2023Article
- Efficient Gene Knockout in Salivary Gland Epithelial Explant Cultures.Journal of dental research · 2023Article
- A pair of primers facing at the double-strand break site enables to detect NHEJ-mediated indel mutations at a 1-bp resolution.Scientific reports · 2022Article
- Histone editing elucidates the functional roles of H3K27 methylation and acetylation in mammals.Nature genetics · 2022Article
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- A Gene Transfer-Positive Cell Sorting System Utilizing Membrane-Anchoring Affinity Tag.Frontiers in bioengineering and biotechnology · 2022Article
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- Functional Screenings Identify Regulatory Variants Associated with Breast Cancer Susceptibility.Current issues in molecular biology · 2021Article
- Parallel Reporter Assays Identify Altered Regulatory Role of rs684232 in Leading to Prostate Cancer Predisposition.International journal of molecular sciences · 2021Article
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- An Editing-Site-Specific PCR Method for Detection and Quantification ofFoods (Basel, Switzerland) · 2021Article
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
CRISPR/Cas9 technology has been widely used for targeted genome modification both in vivo and in vitro. However, an effective method for evaluating genome editing efficiency and screening single-cell clones for desired modification is still lacking. Here, we developed this real time PCR method based on the sensitivity of Taq DNA polymerase to nucleotide mismatch at primer 3' end during initiating DNA replication. Applications to CRISPR gRNAs targeting EMX1, DYRK1A and HOXB13 genes in Lenti-X 293 T cells exhibited comprehensive advantages. Just in one-round qPCR analysis using genomic DNA from cells underwent CRISPR/Cas9 or BE4 treatments, the genome editing efficiency could be determined accurately and quickly, for indel, HDR as well as base editing. When applied to single-cell clone screening, the genotype of each cell colony could also be determined accurately. This method defined a rigorous and practical way in quantify genome editing events.
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