Pairwise Engineering of Tandemly Aligned Self-Splicing Group I Introns for Analysis and Control of Their Alternative Splicing

Author:

Ueda Tomoki1,Nishimura Kei-ichiro2,Nishiyama Yuka1,Tominaga Yuto2,Miyazaki Katsushi1,Furuta Hiroyuki2ORCID,Matsumura Shigeyoshi13,Ikawa Yoshiya13ORCID

Affiliation:

1. Department of Chemistry, Graduate School of Science and Engineering, University of Toyama, Gofuku 3190, Toyama 930-8555, Japan

2. Department of Chemistry and Biochemistry, Graduate School of Engineering, Kyushu University, Moto-oka 744, Nishi-ku, Fukuoka 819-0395, Japan

3. Graduate School of Innovative Life Science, University of Toyama, Gofuku 3190, Toyama 930-8555, Japan

Abstract

Alternative splicing is an important mechanism in the process of eukaryotic nuclear mRNA precursors producing multiple protein products from a single gene. Although group I self-splicing introns usually perform regular splicing, limited examples of alternative splicing have also been reported. The exon-skipping type of splicing has been observed in genes containing two group I introns. To characterize splicing patterns (exon-skipping/exon-inclusion) of tandemly aligned group I introns, we constructed a reporter gene containing two Tetrahymena introns flanking a short exon. To control splicing patterns, we engineered the two introns in a pairwise manner to design pairs of introns that selectively perform either exon-skipping or exon-inclusion splicing. Through pairwise engineering and biochemical characterization, the structural elements important for the induction of exon-skipping splicing were elucidated.

Funder

University of Toyama Discretionary Funds of the President

Publisher

MDPI AG

Subject

Molecular Biology,Biochemistry

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