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Abstract
<p class="first" id="P2">Engineering of the translation apparatus has permitted the
site-specific incorporation
of nonstandard amino acids (nsAAs) into proteins, thereby expanding the genetic code
of organisms. Conventional approaches have focused on porting tRNAs and aminoacyl-tRNA
synthetases (aaRS) from archaea into bacterial and eukaryotic systems where they have
been engineered to site-specifically encode nsAAs. More recent work in genome engineering
has opened up the possibilities of whole genome recoding, in which organisms with
alternative genetic codes have been constructed whereby codons removed from the genetic
code can be repurposed as new sense codons dedicated for incorporation of nsAAs. These
advances, together with the advent of engineered ribosomes and new molecular evolution
methods, enable multisite incorporation of nsAAs and unnatural monomers paving the
way for the template-directed production of functionalized proteins, new classes of
polymers, and genetically encoded materials.
</p>