# Studies of Transfer RNA

> **NIH NIH R35** · YALE UNIVERSITY · 2020 · $16,237

## Abstract

PROJECT SUMMARY
Proteins are typically synthesized with 20 amino acids, yet over 300 amino acids are found in proteins as a result
of posttranslational modifications (PTMs). These natural noncanonical amino acids (ncAAs) modulate protein
function and control fundamental cellular processes. Satisfactory genetic encoding of ncAAs requires the
development of efficient and accurate aminoacyl-tRNA formation and delivery to the ribosome by design of
tRNAs, tRNA synthetases, and elongation factors that constitute orthogonal translation systems (OTSs). While
some ncAAs have been genetically encoded (e.g., N-acetyllysine, phosphoserine (Sep)), OTSs have not been
established for a number of critical PTMs. The overall goal of this proposal is to rewire translation by developing
OTSs for facile and precise production of natural and engineered proteins containing naturally occurring and
synthetic ncAAs.
 These general goals will be realized in three specific areas of the proposed work. (1) Selenium, in the form
of selenocysteine (Sec), is an essential trace element for human health, exhibiting many advantageous chemical
properties with its misincorporation implicated in many disease states. We will engineer efficient site-directed
insertion of Sec and investigate the effects its insertion along with its precursor Sep into several enzymes of
industrial and medical interest. (2) While the genetic code was once thought to be universal, natural codon
reassignments in nature are now known to be widespread. We will couple bioinformatic analysis with our
knowledge of tRNA identity elements to both reveal novel genetic codes and better characterize the role of this
variability in nature. Additionally, we will use long-term evolution to produce an organism with a new genetic code
utilizing synthetic amino acids. (3) We plan to create aminoacyl-tRNA synthetases for efficient synthesis of ncAA-
tRNA for a series of phosphoamino acids and chemically reactive synthetic amino acids. Given the critical role
of phosphorylation in cell signaling and the success of kinase inhibitors against cancer cells, and based on our
success establishing an OTS for phosphoserine, we propose to establish OTSs for additional phosphoamino
acids and their non-hydrolyzable analogs. Incorporation of chemically reactive amino acids will provide a robust
tool to introduce PTMs, biophysical probes, or other valuable residues into a protein of interest.
 The proposed work is significant because the ability to produce, purify, biochemically and structurally char-
acterize proteins containing ncAAs at defined sites is essential for elucidation of fundamental cellular processes
and for construction of new tools for protein design. The innovation of the proposed work is to genetically encode
these biologically relevant ncAAs, and provide efficient OTSs for biochemical and biomedical researchers to help
unravel the complex network of PTMs and their role in biotechnology and human health.

## Key facts

- **NIH application ID:** 10105596
- **Project number:** 3R35GM122560-04S1
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** DIETER SOLL
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $16,237
- **Award type:** 3
- **Project period:** 2017-04-01 → 2022-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10105596

## Citation

> US National Institutes of Health, RePORTER application 10105596, Studies of Transfer RNA (3R35GM122560-04S1). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10105596. Licensed CC0.

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