Hacking the genetic code of mammalian cells.

Area of Science:

• Synthetic biology
• Molecular biology
• Genetics

Background:

• The genetic code dictates protein synthesis.
• Expanding the genetic code allows for the incorporation of novel amino acids.
• Current methods for genetic code expansion in mammalian cells are limited.

Purpose of the Study:

• To develop a straightforward scheme for enlarging the genetic code of mammalian cells.
• To demonstrate the feasibility of transferring an engineered tRNA/aminoacyl-tRNA synthetase pair from E. coli to human cells.
• To incorporate a novel amino acid into proteins in a human cell line.

Main Methods:

• Evolving an orthogonal transfer RNA (tRNA) and aminoacyl-tRNA synthetase (aaRS) pair in E. coli.
• Transferring the evolved orthogonal tRNA/aaRS pair into a human cell line.
• Introducing a photocaged lysine derivative as the novel amino acid.
• Assessing the incorporation of the novel amino acid into cellular proteins.

Main Results:

• Successfully evolved an orthogonal tRNA/aaRS pair in E. coli.
• Demonstrated successful transfer and function of the orthogonal pair in a human cell line.
• Confirmed the incorporation of the photocaged lysine derivative into the human cell's proteome.
• Established a method for enlarging the genetic repertoire of mammalian cells.

Conclusions:

• The developed genetic shuttle system enables the expansion of the genetic code in mammalian cells.
• This approach provides a versatile platform for introducing non-canonical amino acids.
• The methodology holds potential for advancing protein engineering and synthetic biology applications in human cells.