ArticleCellular and molecular life sciences : CMLS2015
Dual-reporter surrogate systems for efficient enrichment of genetically modified cells.
Article in Cellular and molecular life sciences : CMLS, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed, 40 citations in OpenAlex.
- Ultrasound Control of Genomic Regulatory Toolboxes for Cancer Immunotherapy.Nature communications · 2024Article
- Enrichment strategies to enhance genome editing.Journal of biomedical science · 2023Review
- New advances in CRISPR/Cas-mediated precise gene-editing techniques.Disease models & mechanisms · 2023Review
- Modeling long QT syndrome type 2 on-a-chip via in-depth assessment of isogenic gene-edited 3D cardiac tissues.Science advances · 2022Article
- Development of a universal antibiotic resistance screening reporter for improving efficiency of cytosine and adenine base editing.The Journal of biological chemistry · 2022Article
- A high-efficiency and versatile CRISPR/Cas9-mediated HDR-based biallelic editing system.Journal of Zhejiang University. Science. B · 2022Article
- Rapid Assessment of CRISPR Transfection Efficiency and Enrichment of CRISPR Induced Mutations Using a Dual-Fluorescent Stable Reporter System.Frontiers in genome editing · 2022Article
- chi-miR-487b-3p Inhibits Goat Myoblast Proliferation and Differentiation by Targeting IRS1 through the IRS1/PI3K/Akt Signaling Pathway.International journal of molecular sciences · 2021Article
- Comprehensive optimization of a reporter assay toolbox for three distinct CRISPR-Cas systems.FEBS open bio · 2021Article
- Cytosine and adenosine base editing in human pluripotent stem cells using transient reporters for editing enrichment.Nature protocols · 2021Article
- [Rapid screening of single guide RNA targeting pig genome and the harvesting of monoclonal cells by microarray seal].Sheng wu yi xue gong cheng xue za zhi = Journal of biomedical engineering = Shengwu yixue gongchengxue zazhi · 2021Article
- Single-Strand Annealing in Cancer.International journal of molecular sciences · 2021Review
- Strategies for High-Efficiency Mutation Using the CRISPR/Cas System.Frontiers in cell and developmental biology · 2021Review
- A Universal Surrogate Reporter for Efficient Enrichment of CRISPR/Cas9-Mediated Homology-Directed Repair in Mammalian Cells.Molecular therapy. Nucleic acids · 2020Article
- CRISPR/Cas9-facilitated engineering with growth-coupled and sensor-guidedMetabolic engineering communications · 2019Article
- sgRNA-shRNA Structure Mediated SNP Site Editing on PorcineFrontiers in genetics · 2019Article
- Gene cloning and seamless site-directed mutagenesis using single-strand annealing (SSA).Applied microbiology and biotechnology · 2018Article
- dCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression.Nucleic acids research · 2017Article
- An efficient method to enrich for knock-out and knock-in cellular clones using the CRISPR/Cas9 system.Cellular and molecular life sciences : CMLS · 2017Article
- A stable but reversible integrated surrogate reporter for assaying CRISPR/Cas9-stimulated homology-directed repair.The Journal of biological chemistry · 2017Article
Corrections and comments
- Erratum issued
Authors and funding
7 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Isolation of genetically modified cells generated by designed nucleases are challenging, since they are often phenotypically indistinguishable from their parental cells. To efficiently enrich genetically modified cells, we developed two dual-reporter surrogate systems, namely NHEJ-RPG and SSA-RPG based on NHEJ and SSA repair mechanisms, respectively. Repair and enrichment efficiencies of these two systems were compared using different nucleases. In both CRISPR-Cas9- and ZFNs-induced DSB repair studies, we found that the efficiency and sensitivity of the SSA-RPG reporter with direct repeat length more than 200 bp were much higher than the NHEJ-RPG reporter. By utilizing the SSA-RPG reporter, we achieved the enrichment for indels in several endogenous loci with 6.3- to 34.8-fold of non-selected cells. Thus, the highly sensitive SSA-RPG reporter can be used for activity validation of designed nucleases and efficient enrichment of genetically modified cells. Besides, our systems offer alternative enrichment choices either by puromycin selection or FACS.
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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.