Evidence mapPaperPMID 40983722Full record

ReviewNature protocols2026

Selecting aminoacyl-tRNA synthetase/tRNA pairs for efficient genetic encoding of noncanonical amino acids into proteins.

Nathan D Alexander, Yogesh M Gangarde, Riley M Bednar, P Andrew Karplus, Richard B Cooley, Ryan A Mehl

Abstract readReview
In one paragraph

Review in Nature protocols, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

2 citing papers in PubMed.

  1. Review
  2. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

6 authors.

Nathan D Alexander *GCE4All Research Center, Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA.ORCID 0009-0004-2551-6094
Yogesh M Gangarde *GCE4All Research Center, Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA.
Riley M BednarGCE4All Research Center, Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA.
P Andrew KarplusGCE4All Research Center, Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA.
Richard B CooleyGCE4All Research Center, Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA.
Ryan A MehlGCE4All Research Center, Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, USA. ryan.mehl@oregonstate.edu.

Funding

The GCE4All Center: Unleashing the Potential of Genetic Code Expansion for Biomedical ResearchRM1GM144227 · NIGMS · OREGON STATE UNIVERSITY · PI RYAN A MEHL · 2022 to 2026
$6.2M
NIGMS NIH HHS RM1 GM144227U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (NIGMS) RM1-GM144227
6 · The paper itself

Abstract

A critical component of genetic code expansion applications is an aminoacyl-tRNA synthetase (RS)/tRNA pair that faithfully encodes a noncanonical amino acid (ncAA) in response to a specific codon. Here we detail a procedure to select an ncAA-specific RS from a publicly available 3.2-million-member Methanomethylophilus alvus pyrrolysyl-RS (MaPylRS) active site mutant library. Four main parts of the procedure are: (1) preparing the library for use and creating needed cell lines; (2) life and death selections that, respectively, select for functional RSs and select against RSs that incorporate canonical amino acids; (3) three fluorescence-based status checks that provide information about the efficiency and fidelity of the surviving RSs in incorporating the target ncAA; and (4) characterizing top hits to find the best ones for use in applications. The resulting RS/tRNA pairs can be used in either bacterial or eukaryotic cells to study proteins of interest. Additionally, the stability of the MaPylRSs makes them useful in cell-free ncAA-protein expression and amenable to structural and other in vitro characterizations. This Protocol is usable by those with basic molecular biology expertise and features a reliable positive control scheme for selections, status checks at different stages to interpret the level of success and a robust procedure to characterize newly engineered tRNA-RS pairs. Users of this Protocol can expect to select ncAA-specific RS/tRNA pairs from the library within about 30-50 d depending on preparation needs.

Indexed as

Amino AcidsAmino Acyl-tRNA SynthetasesRNA, TransferGene LibraryGenetic CodeProtein EngineeringAmino AcidsAmino Acyl-tRNA SynthetasesRNA, Transfer

Identifiers

PMID40983722
PMCPMC12671338

What Socratic holds

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Registered trials

None linked

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 Socratic graph.