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Nano-optical method for transforming a single yeast cell using exogenous genes.

Yao-Xiong Huang1, Ji-Wang Yang1, Zhuo Wang1

  • 1Department of Biomedical Engineering, Ji Nan University Guangzhou China 510632 tyxhuang@jnu.edu.cn +86-20-85223742 +86-20-85223742.

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Summary

This study introduces a novel nano-optical method for yeast cell transformation using laser tweezers and scissors to deliver exogenous DNA efficiently. The technique achieved a 28% transformation efficiency, enabling stable, multi-generational expression of introduced genes.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Nanotechnology

Background:

  • Efficiently transforming yeast cells with exogenous genes is crucial for various biotechnological applications.
  • Existing methods often face limitations in efficiency, scalability, or genetic material compatibility.

Purpose of the Study:

  • To develop and demonstrate a highly efficient nano-optical method for transforming single yeast cells.
  • To enable direct delivery of naked DNA into yeast cells for stable gene expression.

Main Methods:

  • Utilized laser tweezers/micromanipulators to immobilize yeast cells in a DNA solution.
  • Employed laser scissors to create transient nano-holes in cell walls for DNA delivery.
  • Transformed *Saccharomyces cerevisiae* with Green Fluorescent Protein (GFP) plasmid and bacterial GF1 DNA.

Main Results:

  • Achieved a high transformation efficiency of 28% with a notable survival rate.
  • Recombinant GFP-yeast exhibited stable green fluorescence across multiple generations.
  • Successfully integrated GF1 DNA into the yeast genome, conferring enhanced biomass-digesting capabilities.

Conclusions:

  • The developed nano-optical method offers a highly efficient and versatile approach for yeast genetic transformation.
  • This technique facilitates stable, heritable gene expression, opening avenues for engineered yeast strains with novel functions.
  • The method shows potential for application in various cell types with cell walls for diverse gene transformation needs.