posted on 2024-01-26, 23:07authored byWes Brown, Lance A. Davidson, Alexander Deiters
The
incorporation of unnatural amino acids into proteins
through
genetic code expansion has been successfully adapted to African claw-toed
frog embryos. Six unique unnatural amino acids are incorporated site-specifically
into proteins and demonstrate robust and reliable protein expression.
Of these amino acids, several are caged analogues that can be used
to establish conditional control over enzymatic activity. Using light
or small molecule triggers, we exhibit activation and tunability of
protein functions in live embryos. This approach was then applied
to optical control over the activity of a RASopathy mutant of NRAS,
taking advantage of generating explant cultures from Xenopus. Taken together, genetic code expansion is a robust approach in
the Xenopus model to incorporate novel chemical functionalities
into proteins of interest to study their function and role in a complex
biological setting.