Reprogramming the Cleavage Specificity of Botulinum Neurotoxin Serotype B1.

ABSTRACT: Proteases with reprogrammed specificity for nonnative substrates are highly desired in synthetic biology and biomedicine. However, generating reprogrammed proteases that are orthogonal and highly specific for a new target has been a major challenge. In this work, we sought to expand the versatility of protease systems by engineering an orthogonal botulinum neurotoxin serotype B (BoNT/B) protease that recognizes an orthogonal substrate. We designed and validated an orthogonal BoNT/B protease system in mammalian cells, combining mutations in the protease with compensatory mutations in the protease substrate and incorporating a truncated target sequence and then demonstrated use of this orthogonal BoNT/B protease-substrate combination to regulate complex transcriptional circuitry in mammalian

SUBMITTER: Cleveland JD 

PROVIDER: S-EPMC9907380 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature