title: Short Synthetic Terminators for Improved Heterologous Gene Expression in Yeast.
abstract: Terminators play an important role both in completing the transcription process and impacting mRNA half-life. As such, terminators are an important synthetic component considered in applications such as heterologous gene expression and metabolic engineering. Here, we describe a panel of short (35-70 bp) synthetic terminators that can be used for modulating gene expression in yeast. The best of these synthetic terminator resulted in 3.7-fold more fluorescent protein output and 4.4-fold increase in transcript level compared to that with the commonly used CYC1 terminator. These synthetic terminators offer several advantages over native sequences, including an easily synthesized short length, minimal sequence homology to native sequences, and similar or better performance characteristics than those of commonly used longer terminators. Furthermore, the synthetic terminators are highly functional in both Saccharomyces cerevisiae and an alternative yeast, Yarrowia lipolytica, demonstrating that these synthetic designs are transferrable between diverse yeast species.
authors: †Department of Chemical Engineering, The University of Texas at Austin, 200 East Dean Keeton Street, Stop C0400, Austin, Texas 78712, United States.
authors: Kathleen A
authors: KA
authors: Curran
authors: †Department of Chemical Engineering, The University of Texas at Austin, 200 East Dean Keeton Street, Stop C0400, Austin, Texas 78712, United States.
authors: Nicholas J
authors: NJ
authors: Morse
authors: †Department of Chemical Engineering, The University of Texas at Austin, 200 East Dean Keeton Street, Stop C0400, Austin, Texas 78712, United States.
authors: Kelly A
authors: KA
authors: Markham
authors: †Department of Chemical Engineering, The University of Texas at Austin, 200 East Dean Keeton Street, Stop C0400, Austin, Texas 78712, United States.
authors: Allison M
authors: AM
authors: Wagman
authors: †Department of Chemical Engineering, The University of Texas at Austin, 200 East Dean Keeton Street, Stop C0400, Austin, Texas 78712, United States.
authors: Akash
authors: A
authors: Gupta
authors: †Department of Chemical Engineering, The University of Texas at Austin, 200 East Dean Keeton Street, Stop C0400, Austin, Texas 78712, United States.|‡Institute for Cellular and Molecular Biology, The University of Texas at Austin, 2500 Speedway Avenue, Austin, Texas 78712, United States.
authors: Hal S
authors: HS
authors: Alper
mesh_terms: D020413:3' Untranslated Regions; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D045304:Cytochromes c / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D060847:Metabolic Engineering; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D013728:Terminator Regions, Genetic
chemical_list: D020413:3' Untranslated Regions; D001426:Bacterial Proteins; C014059:CYC1 protein, S cerevisiae; D008164:Luminescent Proteins; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; C054562:yellow fluorescent protein, Bacteria; D045304:Cytochromes c
keywords: 3′ UTR; Saccharomyces cerevisiae; heterologous gene expression; terminator; yeast