ArticleScientific reports2014
An efficient genotyping method for genome-modified animals and human cells generated with CRISPR/Cas9 system.
Article in Scientific reports, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 149 papers.
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149 citing papers in PubMed.
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- Small Nucleolar RNA Snord17 Promotes Self-Renewal of Intestinal Stem Cells through Yy2 mRNA Export and Tead4 Activation.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- Enhancing papaya resistance to ringspot virus through CRISPR/Cas9-mediated gene editing of eFrontiers in plant science · 2026Article
- Bi-allelic deleterious variants in SNAPIN, which encodes a retrograde dynein adaptor, cause a prenatal-onset neurodevelopmental disorder.American journal of human genetics · 2025Article
- Exogenous pyruvate is therapeutic against colitis by targeting cytosolic phospholipase A2.Genes & diseases · 2025Article
- Genomic and biochemical comparison of allelic triple-mutant lines derived from conventional breeding and multiplex gene editing.The plant genome · 2025Article
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- Transcriptome analysis of atad3-null zebrafish embryos elucidates possible disease mechanisms.Orphanet journal of rare diseases · 2025Article
- Detection of CRISPR/Cas9-Mediated Fetal Hemoglobin Reactivation in Erythroblasts Derived from Cord Blood-Hematopoietic Stem Cells.Molecular biotechnology · 2025Article
- RNA methyltransferase SPOUT1/CENP-32 links mitotic spindle organization with the neurodevelopmental disorder SpADMiSS.Nature communications · 2025Article
- Cellular retinoic acid-binding proteins regulate germ cell proliferation and sex determination in zebrafish.Development (Cambridge, England) · 2024Article
- Severe neurodegeneration in brains of transgenic rats producing human tau prions.Acta neuropathologica · 2024Article
- Navigating the CRISPR/Cas Landscape for Enhanced Diagnosis and Treatment of Wilson's Disease.Cells · 2024Review
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- Systematic Comparison of Computational Tools for Sanger Sequencing-Based Genome Editing Analysis.Cells · 2024Article
- Low dose ribosomal DNA P-loop mutation affects development and enforces autophagy in Arabidopsis.RNA biology · 2024Article
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- RNA helicase DDX5 modulates sorafenib sensitivity in hepatocellular carcinoma via the Wnt/β-catenin-ferroptosis axis.Cell death & disease · 2023Article
- Using Ribonucleoprotein-based CRISPR/Cas9 to Edit Single Nucleotide on Human Induced Pluripotent Stem Cells to Model Type 3 Long QT Syndrome (SCN5AStem cell reviews and reports · 2023Article
- Genotyping MUltiplexed-Sequencing of CRISPR-Localized Editing (GMUSCLE): An Experimental and Computational Approach for Analyzing CRISPR-Edited Cells.The CRISPR journal · 2023Article
89 more citing papers are in PubMed but not listed here.
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Authors and funding
14 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The rapid generation of various species and strains of laboratory animals using CRISPR/Cas9 technology has dramatically accelerated the interrogation of gene function in vivo. So far, the dominant approach for genotyping of genome-modified animals has been the T7E1 endonuclease cleavage assay. Here, we present a polyacrylamide gel electrophoresis-based (PAGE) method to genotype mice harboring different types of indel mutations. We developed 6 strains of genome-modified mice using CRISPR/Cas9 system, and utilized this approach to genotype mice from F0 to F2 generation, which included single and multiplexed genome-modified mice. We also determined the maximal detection sensitivity for detecting mosaic DNA using PAGE-based assay as 0.5%. We further applied PAGE-based genotyping approach to detect CRISPR/Cas9-mediated on- and off-target effect in human 293T and induced pluripotent stem cells (iPSCs). Thus, PAGE-based genotyping approach meets the rapidly increasing demand for genotyping of the fast-growing number of genome-modified animals and human cell lines created using CRISPR/Cas9 system or other nuclease systems such as TALEN or ZFN.
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