Rapid evolution of regulatory element libraries for tunable transcriptional and translational control of gene expression

TY - JOUR

T1 - Rapid evolution of regulatory element libraries for tunable transcriptional and translational control of gene expression

AU - Jin, Erqing

AU - Wong, Lynn

AU - Jiao, Yun

AU - Engel, Jake

AU - Holdridge, Benjamin

AU - Xu, Peng

N1 - Publisher Copyright: © 2017 The Authors

PY - 2017/12

Y1 - 2017/12

N2 - Engineering cell factories for producing biofuels and pharmaceuticals has spurred great interests to develop rapid and efficient synthetic biology tools customized for modular pathway engineering. Along the way, combinatorial gene expression control through modification of regulatory element offered tremendous opportunity for fine-tuning gene expression and generating digital-like genetic circuits. In this report, we present an efficient evolutionary approach to build a range of regulatory control elements. The reported method allows for rapid construction of promoter, 5′UTR, terminator and trans-activating RNA libraries. Synthetic overlapping oligos with high portion of degenerate nucleotides flanking the regulatory element could be efficiently assembled to a vector expressing fluorescence reporter. This approach combines high mutation rate of the synthetic DNA with the high assembly efficiency of Gibson Mix. Our constructed library demonstrates broad range of transcriptional or translational gene expression dynamics. Specifically, both the promoter library and 5′UTR library exhibits gene expression dynamics spanning across three order of magnitude. The terminator library and trans-activating RNA library displays relatively narrowed gene expression pattern. The reported study provides a versatile toolbox for rapidly constructing a large family of prokaryotic regulatory elements. These libraries also facilitate the implementation of combinatorial pathway engineering principles and the engineering of more efficient microbial cell factory for various biomanufacturing applications.

AB - Engineering cell factories for producing biofuels and pharmaceuticals has spurred great interests to develop rapid and efficient synthetic biology tools customized for modular pathway engineering. Along the way, combinatorial gene expression control through modification of regulatory element offered tremendous opportunity for fine-tuning gene expression and generating digital-like genetic circuits. In this report, we present an efficient evolutionary approach to build a range of regulatory control elements. The reported method allows for rapid construction of promoter, 5′UTR, terminator and trans-activating RNA libraries. Synthetic overlapping oligos with high portion of degenerate nucleotides flanking the regulatory element could be efficiently assembled to a vector expressing fluorescence reporter. This approach combines high mutation rate of the synthetic DNA with the high assembly efficiency of Gibson Mix. Our constructed library demonstrates broad range of transcriptional or translational gene expression dynamics. Specifically, both the promoter library and 5′UTR library exhibits gene expression dynamics spanning across three order of magnitude. The terminator library and trans-activating RNA library displays relatively narrowed gene expression pattern. The reported study provides a versatile toolbox for rapidly constructing a large family of prokaryotic regulatory elements. These libraries also facilitate the implementation of combinatorial pathway engineering principles and the engineering of more efficient microbial cell factory for various biomanufacturing applications.

KW - 5′UTR

KW - Library

KW - Promoter

KW - Riboregulatory

KW - Synthetic biology

KW - Terminator

KW - Trans-activating RNA

UR - http://www.scopus.com/inward/record.url?scp=85047209553&partnerID=8YFLogxK

U2 - 10.1016/j.synbio.2017.10.003

DO - 10.1016/j.synbio.2017.10.003

M3 - 文章

AN - SCOPUS:85047209553

SN - 2405-805X

VL - 2

SP - 295

EP - 301

JO - Synthetic and Systems Biotechnology

JF - Synthetic and Systems Biotechnology

IS - 4

ER -