ArticleCellular and molecular life sciences : CMLS2019
LION: a simple and rapid method to achieve CRISPR gene editing.
Article in Cellular and molecular life sciences : CMLS, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed, 5 citations in OpenAlex.
- Prospects and challenges in using engineered lactic acid bacteria in aquaculture applications.Engineering microbiology · 2026Review
- Systematical analysis reveals a strong cancer relevance of CREB1-regulated genes.Cancer cell international · 2021Article
- Efficient correction of Duchenne muscular dystrophy mutations by SpCas9 and dual gRNAs.Molecular therapy. Nucleic acids · 2021Article
- Haplotyping by CRISPR-mediated DNA circularization (CRISPR-hapC) broadens allele-specific gene editing.Nucleic acids research · 2020Article
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Authors and funding
19 authors at 6 institutions in 3 countries.
Funding
Abstract
The RNA-guided CRISPR-Cas9 technology has paved the way for rapid and cost-effective gene editing. However, there is still a great need for effective methods for rapid generation and validation of CRISPR/Cas9 gRNAs. Previously, we have demonstrated that highly efficient generation of multiplexed CRISPR guide RNA (gRNA) expression array can be achieved with Golden Gate Assembly (GGA). Here, we present an optimized and rapid method for generation and validation in less than 1 day of CRISPR gene targeting vectors. The method (LION) is based on ligation of double-stranded gRNA oligos into CRISPR vectors with GGA followed by nucleic acid purification. Using a dual-fluorescent reporter vector (C-Check), T7E1 assay, TIDE assay and a traffic light reporter assay, we proved that the LION-based generation of CRISPR vectors are functionally active, and equivalent to CRISPR plasmids generated by traditional methods. We also tested the activity of LION CRISPR vectors in different human cell types. The LION method presented here advances the rapid functional validation and application of CRISPR system for gene editing and simplified the CRISPR gene-editing procedures.
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Registered trials
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