Work

High Yield, Low Magnesium Flexizyme Reactions in a Water-Ice Eutectic Phase

Year: 2025

Type: article

Abstract: Flexizymes enable the stoichiometric acylation of tRNAs with a variety of compounds, enabling the in vitro translation of peptides with both non-natural backbones and side chains. However, flexizyme reactions have several drawbacks, including single-turnover kinetics, high Mg(II) carryover inhibiting in vitro translation, and rapid product hydrolysis. Here we present flexizyme reactions utilizing an ice-eutectic phase, with high yields, 30X lower Mg(II), and long-term product stability. The eutectic flexizyme reactions increase the ease of use and flexibility of flexizyme aminoacylation, and increase the in vitro protein production.

Source: Biochemistry

DOI: 10.1021/acs.biochem.5c00371

Authors: Joshua A. Davisson, Jose L. Alejo, Mace Blank, Evan M. Kalb, Angelin Prasad, Isaac J. Knudson, Alanna Schepartz, Aaron E. Engelhart, Katarzyna P. Adamala

Institutions: University of Minnesota, University of Minnesota System, University of California, Berkeley, St. Jude Children's Research Hospital

Language: English

Topic: RNA and protein synthesis mechanisms

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: National Science Foundation, Howard Hughes Medical Institute, Alfred P. Sloan Foundation

Awards: CAREER: Nucleic Acid-Catalyzed Halogenations, G3792488682, NSF Center for Genetically Encoded Materials, Collaborative Research: Using non-canonical genetic alphabets to expand genetic codes in synthetic cells, GOALI: SemiSynBio-III: Moving Millions of Droplets at Megahertz Speeds: DNA Computing, DNA Storage, and Synthetic Biology on an Industrial Platform for Digital Microfluidics, Research - University of Minnesota (2024)

OpenAlex ID: W4413353263

FWCI: 0.52

Cites: 23

Cited by: 2

Related to: 10