Programmable mutually exclusive alternative splicing for generating RNA and protein diversity.

Published

Journal Article

Alternative splicing performs a central role in expanding genomic coding capacity and proteomic diversity. However, programming of splicing patterns in engineered biological systems remains underused. Synthetic approaches thus far have predominantly focused on controlling expression of a single protein through alternative splicing. Here, we describe a modular and extensible platform for regulating four programmable exons that undergo a mutually exclusive alternative splicing event to generate multiple functionally-distinct proteins. We present an intron framework that enforces the mutual exclusivity of two internal exons and demonstrate a graded series of consensus sequence elements of varying strengths that set the ratio of two mutually exclusive isoforms. We apply this framework to program the DNA-binding domains of modular transcription factors to differentially control downstream gene activation. This splicing platform advances an approach for generating diverse isoforms and can ultimately be applied to program modular proteins and increase coding capacity of synthetic biological systems.

Full Text

Duke Authors

Cited Authors

  • Mathur, M; Kim, CM; Munro, SA; Rudina, SS; Sawyer, EM; Smolke, CD

Published Date

  • June 17, 2019

Published In

Volume / Issue

  • 10 / 1

Start / End Page

  • 2673 -

PubMed ID

  • 31209208

Pubmed Central ID

  • 31209208

Electronic International Standard Serial Number (EISSN)

  • 2041-1723

International Standard Serial Number (ISSN)

  • 2041-1723

Digital Object Identifier (DOI)

  • 10.1038/s41467-019-10403-w

Language

  • eng