Interrogating the Function of Bicistronic Translational Control Elements to Improve Consistency of Gene Expression

Context independent gene expression is required for genetic circuits to maintain consistent and predicable behavior. Previous efforts to develop context independent translation have leveraged the helicase activity of translating ribosomes via bicistronic design translational control elements (BCDs)...

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Bibliographic Details
Published in:	ACS synthetic biology Vol. 12; no. 6; pp. 1608 - 1615
Main Authors:	Jansen, Zachary, Reilly, Sophia R., Lieber-Kotz, Matan, Li, Andrew Z., Wei, Qiyao, Kulhanek, Devon L., Gilmour, Andrew R., Thyer, Ross
Format:	Journal Article
Language:	English
Published:	United States American Chemical Society 16-06-2023
Subjects:	translation synthetic biology Rhodococcus ribosome binding sites golden gate assembly nonmodel bacteria
Online Access:	Get full text
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Summary:	Context independent gene expression is required for genetic circuits to maintain consistent and predicable behavior. Previous efforts to develop context independent translation have leveraged the helicase activity of translating ribosomes via bicistronic design translational control elements (BCDs) located within an efficiently translated leader peptide. We have developed a series of bicistronic translational control elements with strengths that span several orders of magnitude, maintain consistent expression levels across diverse sequence contexts, and are agnostic to common ligation sequences used in modular cloning systems. We have used this series of BCDs to investigate several features of this design, including the spacing of the start and stop codons, the nucleotide identity upstream of the start codon, and factors affecting translation of the leader peptide. To demonstrate the flexibility of this architecture and their value as a generic modular expression control cassette for synthetic biology, we have developed a set of robust BCDs for use in several Rhodococcus species.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	2161-5063 2161-5063
DOI:	10.1021/acssynbio.3c00093