Diversity-generating retroelements for programmable targeted hypermutagenesis - PubMed
5 hours ago
- #directed evolution
- #hypermutagenesis
- #genetic engineering
- DGRs are natural systems that accelerate evolution through targeted hypermutation.
- DGRec method combines DGRs with recombineering to diversify sequences in E. coli.
- Reverse transcriptase biases maximize sequence exploration while avoiding nonsense mutations.
- DGRec diversifies 50-200 bp windows with high error rates at adenines.
- Mutation rates up to 1.38 × 10^-2 per base per generation allow up to 24 mutations in 48 hours.
- Applications include phage λ host-range engineering, dCas9 evolution, and nanobody optimization.
- DGR-mediated mutagenesis is feasible in yeast using adapted recombination and selection strategies.