GGAssembler: Precise and economical design and synthesis of combinatorial mutation libraries.

ABSTRACT: Golden Gate assembly (GGA) can seamlessly generate full-length genes from DNA fragments. In principle, GGA could be used to design combinatorial mutation libraries for protein engineering, but creating accurate, complex, and cost-effective libraries has been challenging. We present GGAssembler, a graph-theoretical method for economical design of DNA fragments that assemble a combinatorial library that encodes any desired diversity. We used GGAssembler for one-pot in vitro assembly of camelid antibody libraries comprising >10⁵ variants with DNA costs <0.007$ per variant and dropping significantly with increased library complexity. >93% of the desired variants were present in the assembly product and >99% were represented within the expected order of magnitude as verified by deep

SUBMITTER: Hoch SY 

PROVIDER: S-EPMC11403590 | biostudies-literature | 2024 Oct

REPOSITORIES: biostudies-literature