Synthetic Translational Regulation by Protein-Binding RNA Origami Scaffolds

TY - JOUR

T1 - Synthetic Translational Regulation by Protein-Binding RNA Origami Scaffolds

AU - Nguyen, Michael T A

AU - Pothoulakis, Georgios

AU - Andersen, Ebbe S

PY - 2022/5/20

Y1 - 2022/5/20

N2 - Rational design approaches for the regulation of gene expression are expanding the synthetic biology toolbox. However, only a few tools for regulating gene expression at the translational level have been developed. Here, we devise an approach for translational regulation using the MS2 and PP7 aptamer and coat-protein pairs in Escherichia coli. The aptamers are used as operators in transcription units that encode proteins fused to their cognate coat proteins, which leads to self-repression. RNA origami scaffolds that contain up to four aptamers serve as an alternate binder to activate translation. With this system, we demonstrate that the increase in expression of a reporter protein is dependent on both the concentration and number of aptamers on RNA origami scaffolds. We also demonstrate regulation of multiple proteins using a single MS2 coat protein fusion and apply this method to regulate the relative expression of enzymes of the branched pathway for deoxyviolacein biosynthesis.

AB - Rational design approaches for the regulation of gene expression are expanding the synthetic biology toolbox. However, only a few tools for regulating gene expression at the translational level have been developed. Here, we devise an approach for translational regulation using the MS2 and PP7 aptamer and coat-protein pairs in Escherichia coli. The aptamers are used as operators in transcription units that encode proteins fused to their cognate coat proteins, which leads to self-repression. RNA origami scaffolds that contain up to four aptamers serve as an alternate binder to activate translation. With this system, we demonstrate that the increase in expression of a reporter protein is dependent on both the concentration and number of aptamers on RNA origami scaffolds. We also demonstrate regulation of multiple proteins using a single MS2 coat protein fusion and apply this method to regulate the relative expression of enzymes of the branched pathway for deoxyviolacein biosynthesis.

KW - RNA origami

KW - RNA synthetic biology

KW - RNA-protein complexes

KW - post-transcriptional regulation

U2 - 10.1021/acssynbio.1c00608

DO - 10.1021/acssynbio.1c00608

M3 - Letter

C2 - 35438978

SN - 2161-5063

VL - 11

SP - 1710

EP - 1718

JO - ACS Synthetic Biology

JF - ACS Synthetic Biology

IS - 5

ER -