High-throughput recovery of integron cassettes for gene discovery screens.

Cassette gatherer and cassette hunter recover hundreds of single integron cassettes per assay with over 99% specificity in Vibrio cholerae and DNA-derived libraries.

• Why it matters: Integron cassettes are a largely untapped reservoir of functional genes, but existing recovery approaches limit high-throughput, sequence- and function-independent discovery.
• What they did: The authors re-engineered a class 1 integron into counterselection markers on a plasmid or chromosome in naturally competent Vibrio cholerae, then used cassette gatherer/hunter to build gene libraries from tractable strains or directly from DNA.
• The result: Screening the recovered cassette libraries against phages ICP2 and T4 identified nine phage-defence systems, including five previously undescribed, enabling rapid functional gene discovery.

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