Context and number of noncanonical repeat variable diresidues impede the design of TALE proteins with improved DNA targeting.

ABSTRACT: Transcription activator-like effector (TALE) proteins have been used extensively for targeted binding of fusion proteins to loci of interest in (epi)genome engineering. Such approaches typically utilize four canonical TALE repeat variable diresidue (RVD) types, corresponding to the identities of two key amino acids, to target each nucleotide. Alternate RVDs with improved specificity are desired. Here, we focused on seven noncanonical RVDs that have been suggested to have improved specificity for their target nucleotides. We used custom protein binding microarrays to characterize the DNA-binding activity of 65 TALEs containing these alternate or corresponding canonical RVDs at multiple positions to ~5,000 unique DNA sequences per protein. We found that none of the noncanonical thymine-targe

SUBMITTER: Anderson JT 

PROVIDER: S-EPMC6954735 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature