Why tRNA
Transfer RNAs (tRNAs) are key to the ribosomal readout of the genetic code. tRNA shape, stability, amino-acylation, and decoding in translation, all depend on complex post-transcriptional modifications.
High quality tRNA are crucial to all in vitro translation systems, including whole cell lysate and PURE system.
Here we share toolbox of our tRNA Foundry: methods to purify whole tRNA pools from various organisms, and methods to purify single tRNA species.
Our protocols yield tRNA with full complement of post-transcriptional modifications.
tRNA pools - total tRNA
For a long time, our community relied on MRE600 tRNA sold by Roche. Since this product was discontinued, there is an urgent need for a source of a complete tRNA pool for cell-free translation.
Here text from https://docs.google.com/document/d/120NMvEN9l48PkyItzfZa82KaQJqXxG13GL-1dzfeZUA/edit?tab=t.0
Pick and choose: single tRNA species
Many applications can banafit from the ability to tailor custom tRNA supply, including genetic code expansion, codon bias studies and other efforts to engineer modified translation systems.
Here text from https://docs.google.com/document/d/19xn92G-Z4ufYkSCcL8VjbT4LWN3_Maz9pjAe8h8AfdU/edit?tab=t.0
The team
The tRNA project is work in progresss in the Adamala lab, many people are engineering and improving on existing protocols and bringing in new ideas.
The project is led by Evan Kalb, Joshua Davisson and Elisabeth Edgerton.
References
tRNA pools:
tRNA Abundance Determines Translation Yield in Cell-Free Expression Systems;
Evan M. Kalb, Russel M. Vincent, Aaron E. Engelhart, George M. Church, Katarzyna Adamala
coming soon
Single tRNAs:
Purification of post-transcriptionally modified tRNAs for enhanced cell-free translation systems;
Evan M. Kalb, Jose L. Alejo, Leticia Dias-Fields, Isaac Knudson, Joshua A. Davisson, Efren Maldonado, Kanokporn Chattrakun, Shangsi Lin, Alanna Schepartz, Shenglong Zhang, Scott Blanchard, Aaron E. Engelhart, Katarzyna P. Adamala
preprint
https://doi.org/10.1101/2025.06.10.658963