Overview
- The method measures both insertion efficiency and on-target specificity in a single high-throughput experiment.
- The team focused on the V-K CAST subtype and systematically screened a library covering every possible single-point mutation.
- Combining select beneficial mutations increased activity about fivefold while preserving or improving specificity.
- The work, published September 23, 2025 in Nucleic Acids Research, offers mechanistic insight and a platform for iterative design.
- Researchers note that CASTs can integrate large DNA payloads but underperform in human cells, so translation beyond bacterial systems remains a next step.