High-throughput retrieval of target sequences from complex clone libraries using CRISPRi.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 36411313.
- Also identified by DOI 10.1038/s41587-022-01531-8 and PMC identifier 11042918.
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Abstract
The capture of metagenomic DNA in large clone libraries provides the opportunity to study microbial diversity that is inaccessible using culture-dependent methods. In this study, we harnessed nuclease-deficient Cas9 to establish a CRISPR counter-selection interruption circuit (CCIC) that can be used to retrieve target clones from complex libraries. Combining modern sequencing methods with CCIC cloning allows for rapid physical access to the genetic diversity present in natural ecosystems.
Medical subject headings
- Ecosystem
- Metagenomics