Multiplex base editing to convert TAG into TAA codons in the human genome.

Area of Science:

• Synthetic Biology
• Genetics
• Molecular Biology

Background:

• Whole-genome recoding offers advanced capabilities like nonstandard amino acids and enhanced resistance in bacteria.
• Extending these genetic engineering techniques to human cells presents significant opportunities for therapeutic and research applications.

Purpose of the Study:

• To investigate the feasibility of whole-genome recoding in human cells.
• To demonstrate precise base editing for essential genes and assess genome-wide off-target effects.
• To introduce a computational tool for recoding mammalian genomes.

Main Methods:

• Utilized base editing to convert TAG codons to TAA across 33 essential genes in human cells via single transfection.
• Performed genome-wide analysis to identify and quantify off-target editing events.
• Developed and implemented GRIT, a computational tool for genome recoding.

Main Results:

• Achieved exceptional base editing efficiency for essential genes in human cells.
• Observed approximately 40 C-to-T off-target events within essential gene exons during genome-wide analysis.
• Successfully demonstrated the feasibility of multiplex genome editing and recoding in mammalian cells.

Conclusions:

• Whole-genome recoding is feasible in human cells, enabling precise genetic modifications.
• The study highlights the potential for highly multiplexed genome editing in mammalian systems.
• The GRIT tool facilitates computational approaches to genome recoding.