Efficient and Precise Protein Synthesis in a Cell-Free System Using a Set of In Vitro Transcribed tRNAs with Nucleotide Modifications.

Kazuaki Amikura, Keita Hibi, Yoshihiro Shimizu

Abstract read

In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it

0cells of the map it votes in

1citing papers in PubMed

0.1field-weighted citation impact, top 60% of its field

1 · What the graph read from it

What it found

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3 · Its place in the literature

Who cites it

1 citing paper in PubMed, 1 citations in OpenAlex.

Optimizing Human Cell-Free System for Efficient Protein Production.Journal of microbiology and biotechnology · 2025
Article

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5 · Who and what money

Authors and funding

3 authors at 2 institutions in 2 countries.

Kazuaki AmikuraDepartment of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA.

Keita HibiDepartment of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Chiba, Japan.

Yoshihiro ShimizuLaboratory for Cell-Free Protein Synthesis, RIKEN Center for Biosystems Dynamics research (BDR), Osaka, Japan. yshimizu@riken.jp.

The University of Tokyo · JPYale University · US

Funding

Studies of Transfer RNAR35GM122560 · NIGMS · YALE UNIVERSITY · PI SOLL, DIETER · 2017 to 2022

$5.6M

NIGMS NIH HHS R35 GM122560

6 · The paper itself

Abstract

Reconstitution of a complicated system with a minimal set of components is essential for understanding the mechanisms of how the input is reflected in the output, which is fundamental for further engineering of the corresponding system. We have recently developed a reconstituted cell-free protein synthesis system equipped only with 21 in vitro transcribed tRNAs, one of the minimal systems for understanding the genetic code decoding mechanisms. Introduction of several nucleotide modifications to the transcribed tRNAs showed improvement of both protein synthesis efficiency and its fidelity, suggesting various combinations of tRNAs and their modifications can be evaluated in the developed system. In this chapter, we describe how to prepare this minimal system. Methods for preparing the transcribed tRNAs, their modifications, and the protein production using the set of prepared tRNAs are shown.

Indexed as

NucleotidesRNA, TransferCell-Free SystemGenetic CodeProtein BiosynthesisNucleotidesRNA, TransferCell-free protein synthesis systemGenetic codeIn vitro transcriptionNucleotide modificationPURE systemSynthetic biologytRNA

Identifiers

PMID34985743

DOI10.1101/cshperspect.a023812

OpenAlexW4205865639

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.