Programmable Mammalian Translational Modulators by CRISPR-associated Proteins

Overview

Journal Nat Commun

Specialty Biology

Date 2023 Apr 19

PMID 37076490

Authors

Shunsuke Kawasaki

Hiroki Ono

Moe Hirosawa

Takeru Kuwabara

Shunsuke Sumi

Suji Lee

Knut Woltjen

Hirohide Saito

Affiliations

Soon will be listed here.

Abstract

Translational modulation based on RNA-binding proteins can be used to construct artificial gene circuits, but RNA-binding proteins capable of regulating translation efficiently and orthogonally remain scarce. Here we report CARTRIDGE (Cas-Responsive Translational Regulation Integratable into Diverse Gene control) to repurpose Cas proteins as translational modulators in mammalian cells. We demonstrate that a set of Cas proteins efficiently and orthogonally repress or activate the translation of designed mRNAs that contain a Cas-binding RNA motif in the 5'-UTR. By linking multiple Cas-mediated translational modulators, we designed and built artificial circuits like logic gates, cascades, and half-subtractor circuits. Moreover, we show that various CRISPR-related technologies like anti-CRISPR and split-Cas9 platforms could be similarly repurposed to control translation. Coupling Cas-mediated translational and transcriptional regulation enhanced the complexity of synthetic circuits built by only introducing a few additional elements. Collectively, CARTRIDGE has enormous potential as a versatile molecular toolkit for mammalian synthetic biology.

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