Work

In Vitro Transcriptional Regulation via Nucleic-Acid-Based Transcription Factors

Year: 2019

Type: article

Abstract: Cells execute complex transcriptional programs by deploying distinct protein regulatory assemblies that interact with cis-regulatory elements throughout the genome. Using concepts from DNA nanotechnology, we synthetically recapitulated this feature in cell-free gene networks actuated by T7 RNA polymerase (RNAP). Our approach involves engineering nucleic-acid hybridization interactions between a T7 RNAP site-specifically functionalized with single-stranded DNA (ssDNA), templates displaying cis-regulatory ssDNA domains, and auxiliary nucleic-acid assemblies acting as artificial transcription factors (TFs). By relying on nucleic-acid hybridization, de novo regulatory assemblies can be computationally designed to emulate features of protein-based TFs, such as cooperativity and combinatorial binding, while offering unique advantages such as programmability, chemical stability, and scalability. We illustrate the use of nucleic-acid TFs to implement transcriptional logic, cascading, feedback, and multiplexing. This framework will enable rapid prototyping of increasingly complex in vitro genetic devices for applications such as portable diagnostics, bio-analysis, and the design of adaptive materials.

Source: ACS Synthetic Biology

DOI: 10.1021/acssynbio.9b00242

Authors: Leo Y. T. Chou, William M. Shih

Institutions: Harvard University, Dana-Farber Cancer Institute, Wyss Institute for Biologically Inspired Engineering

Language: English

Topic: Advanced biosensing and bioanalysis techniques

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: National Science Foundation, Hansjörg Wyss Institute for Biologically Inspired Engineering, Harvard University

Award: Collaborative Research: Molecular Programming Architectures, Abstractions, Algorithms, and Applications