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Nat Commun. 2019 Jun 17;10(1):2673. doi: 10.1038/s41467-019-10403-w.

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

Author information

1
Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA.
2
Joint Initiative for Metrology in Biology, Stanford, CA, 94305, USA.
3
Genome-scale Measurements Group, National Institute of Standards and Technology, Stanford, CA, 94305, USA.
4
Minnesota Supercomputing Institute, University of Minnesota, Minneapolis, MN, 55455, USA.
5
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, 94720, USA.
6
Department of Bioengineering, Stanford University, Stanford, CA, 94305, USA. csmolke@stanford.edu.
7
Chan Zuckerberg Biohub, San Francisco, CA, 94158, USA. csmolke@stanford.edu.

Abstract

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.

PMID:
31209208
PMCID:
PMC6572816
DOI:
10.1038/s41467-019-10403-w
[Indexed for MEDLINE]
Free PMC Article

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