Advances in in vitro genetic code reprogramming in 2014-2017.

Area of Science:

• Biochemistry
• Molecular Biology
• Synthetic Biology

Background:

• Genetic code manipulation allows introducing non-proteinogenic amino acids into peptides via translation.
• Current methods have limitations in the number of simultaneously usable amino acids and substrate compatibility.
• Difficult amino acid substrates (e.g., d-amino acids, β-amino acids) are inefficiently incorporated compared to l-α-amino acids.

Purpose of the Study:

• To address limitations in the number and types of amino acids incorporated into peptides.
• To explore novel methodologies for expanding the genetic code and improving translation efficiency.
• To enhance the incorporation of challenging amino acid substrates into peptides.

Main Methods:

• Artificial division of codon boxes to expand the number of available codons.
• Development of orthogonal ribosomal translation systems to duplicate the codon table.
• Engineering novel artificial tRNAs to enhance the incorporation of difficult amino acid substrates.

Main Results:

• Demonstrated methodologies to increase the diversity of amino acids in peptides.
• Showcased systems capable of utilizing a broader range of amino acid substrates.
• Improved efficiency of incorporating non-standard amino acids, including d- and β-amino acids.

Conclusions:

• Recent advancements significantly expand the possibilities for incorporating non-proteinogenic amino acids into peptides.
• These methods overcome previous limitations in simultaneous amino acid usage and substrate compatibility.
• The developed strategies offer powerful tools for peptide engineering and synthetic biology applications.