ArticleNucleic acids research2021
A simple and rapid method for enzymatic synthesis of CRISPR-Cas9 sgRNA libraries.
Article in Nucleic acids research, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- RNA chemistry and therapeutics.Nature reviews. Drug discovery · 2025Review
- The type IIS restriction enzyme MmeI can cut across a double-strand break.Molecular biology reports · 2023Article
- A new method to synthesize multiple gRNA libraries and functional mapping of mammalian H3K4me3 regions.Nucleic acids research · 2023Article
- New Target Gene Screening Using Shortened and Random sgRNA Libraries in Microbial CRISPR Interference.ACS synthetic biology · 2023Article
- CRISPR-Cas9 library screening approach for anti-cancer drug discovery: overview and perspectives.Theranostics · 2022Review
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Authors and funding
5 authors.
Funding
Abstract
CRISPR-Cas9 sgRNA libraries have transformed functional genetic screening and have enabled several innovative methods that rely on simultaneously targeting numerous genetic loci. Such libraries could be used in a vast number of biological systems and in the development of new technologies, but library generation is hindered by the cost, time, and sequence data required for sgRNA library synthesis. Here, we describe a rapid enzymatic method for generating robust, variant-matched libraries from any source of cDNA in under 3 h. This method, which we have named SLALOM, utilizes a custom sgRNA scaffold sequence and a novel method for detaching oligonucleotides from solid supports by a strand displacing polymerase. With this method, we constructed libraries targeting the E. coli genome and the transcriptome of developing zebrafish hearts, demonstrating its ability to expand the reach of CRISPR technology and facilitate methods requiring custom libraries.
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Registered trials
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