A modular approach for dCas9-mediated enzyme cascading via orthogonal bioconjugation. Issue 77 (25th August 2020)

• Record Type: Journal Article
• Authors: Berckman, Emily A.; Chen, Wilfred
• Is Part Of: Chemical communications. Volume 56:Issue 77(2020)
• Page Start: 11426
• Page End: 11428
• Publication Date: 2020-08-25
• Subjects: Chemistry -- Periodicals; 540
• DOI: 10.1039/d0cc04196c
• Languages: English
• ISSNs: 1359-7345

Abstract

Abstract : We report a new modular strategy to assemble dCas9-guided enzyme cascades by employing orthogonal post-translation chemistry to enable the one-pot bioconjugation and guided assembly of two different dCas9 enzymes onto a DNA scaffold. Abstract : We report a new modular strategy to assemble dCas9-guided enzyme cascades by employing orthogonal post-translation chemistry. Two orthogonal SpyCatcher and SnoopCatcher pairs were used for the one-pot enzyme bioconjugation onto two different dCas9 proteins to enable their guided assembly onto a DNA scaffold. The resulting two-component cellulosomes exhibited 2.8-fold higher reducing sugar production over unassembled enzymes. This platform retains the high binding affinity afforded by dCas9 proteins for easy control over enzyme assembly while offering the flexibility for both in vivo and in vitro assembly of a wide array of enzyme cascades with minimal optimization.