ArticleCellular and molecular life sciences : CMLS2016
Enhanced genome editing in mammalian cells with a modified dual-fluorescent surrogate system.
Article in Cellular and molecular life sciences : CMLS, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers, 1 of them a synthesis that pooled it.
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Who cites it
21 citing papers in PubMed, 1 synthesis or guideline pooled it.
- An overview and metanalysis of machine and deep learning-based CRISPR gRNA design tools.RNA biology · 2020Pooled it
- Targeted nanodelivery strategies for atrial fibrillation: concomitant targeting of fibrosis suppression and electrical conduction restoration through advanced nanobiotechnology.Journal of nanobiotechnology · 2025Review
- A bibliometric mapping of advancements and trends in genome editing in pigs.Tropical animal health and production · 2025Review
- AMBRA1 phosphorylation by CDK1 and PLK1 regulates mitotic spindle orientation.Cellular and molecular life sciences : CMLS · 2023Article
- Enrichment strategies to enhance genome editing.Journal of biomedical science · 2023Review
- Comparison of In-Frame Deletion, Homology-Directed Repair, and Prime Editing-Based Correction of Duchenne Muscular Dystrophy Mutations.Biomolecules · 2023Article
- A genetically modified minipig model for Alzheimer's disease with SORL1 haploinsufficiency.Cell reports. Medicine · 2022Article
- Generation of Monoclonal iPSC Lines with Stable Cas9 Expression and High Cas9 Activity.Methods in molecular biology (Clifton, N.J.) · 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
- Systematical analysis reveals a strong cancer relevance of CREB1-regulated genes.Cancer cell international · 2021Article
- Haplotyping by CRISPR-mediated DNA circularization (CRISPR-hapC) broadens allele-specific gene editing.Nucleic acids research · 2020Article
- LION: a simple and rapid method to achieve CRISPR gene editing.Cellular and molecular life sciences : CMLS · 2019Article
- Generation and characterization of a novel knockin minipig model of Hutchinson-Gilford progeria syndrome.Cell discovery · 2019Article
- BE-FLARE: a fluorescent reporter of base editing activity reveals editing characteristics of APOBEC3A and APOBEC3B.BMC biology · 2018Article
- CRISPR-C: circularization of genes and chromosome by CRISPR in human cells.Nucleic acids research · 2018Article
- A Modified Monomeric Red Fluorescent Protein Reporter for Assessing CRISPR Activity.Frontiers in cell and developmental biology · 2018Article
- Ligase IV inhibitor SCR7 enhances gene editing directed by CRISPR-Cas9 and ssODN in human cancer cells.Cell & bioscience · 2018Article
- 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
- IGF1R depletion facilitates MET-amplification as mechanism of acquired resistance to erlotinib in HCC827 NSCLC cells.Oncotarget · 2017Article
- CRISPR-mediated multiplexed genetic manipulation.Oncotarget · 2016Article
Corrections and comments
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Authors and funding
18 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Programmable DNA nucleases such as TALENs and CRISPR/Cas9 are emerging as powerful tools for genome editing. Dual-fluorescent surrogate systems have been demonstrated by several studies to recapitulate DNA nuclease activity and enrich for genetically edited cells. In this study, we created a single-strand annealing-directed, dual-fluorescent surrogate reporter system, referred to as C-Check. We opted for the Golden Gate Cloning strategy to simplify C-Check construction. To demonstrate the utility of the C-Check system, we used the C-Check in combination with TALENs or CRISPR/Cas9 in different scenarios of gene editing experiments. First, we disrupted the endogenous pIAPP gene (3.0 % efficiency) by C-Check-validated TALENs in primary porcine fibroblasts (PPFs). Next, we achieved gene-editing efficiencies of 9.0-20.3 and 4.9 % when performing single- and double-gene targeting (MAPT and SORL1), respectively, in PPFs using C-Check-validated CRISPR/Cas9 vectors. Third, fluorescent tagging of endogenous genes (MYH6 and COL2A1, up to 10.0 % frequency) was achieved in human fibroblasts with C-Check-validated CRISPR/Cas9 vectors. We further demonstrated that the C-Check system could be applied to enrich for IGF1R null HEK293T cells and CBX5 null MCF-7 cells with frequencies of nearly 100.0 and 86.9 %, respectively. Most importantly, we further showed that the C-Check system is compatible with multiplexing and for studying CRISPR/Cas9 sgRNA specificity. The C-Check system may serve as an alternative dual-fluorescent surrogate tool for measuring DNA nuclease activity and enrichment of gene-edited cells, and may thereby aid in streamlining programmable DNA nuclease-mediated genome editing and biological research.
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