Novel Microbial Modification Tools to Convert Lipids into Other Value-Added Products

Area of Science:

• Molecular Biology
• Genome Engineering
• Microbial Genetics

Background:

• CRISPR-Cas9 is a microbial adaptive immune system that has revolutionized molecular biology.
• The Type II CRISPR system uses Cas9 nuclease and guide RNA to target specific DNA sequences.
• CRISPR-Cas9 technology enables precise DNA editing and gene expression regulation.

Purpose of the Study:

• To describe a method for CRISPR-Cas9-mediated genome editing in yeast.
• To enable biotechnological applications through precise genetic modifications in yeast.
• To detail a rapid and efficient genome editing procedure in a model organism.

Main Methods:

• CRISPR-Cas9-mediated genome editing utilizing nonhomologous end joining (NHEJ) or homology-directed repair (HDR) pathways.
• Selection of target sequences and cloning of guide RNA (gRNA) with the target sequence.
• Transformation of yeast cells and verification of desired mutations, deletions, or insertions.

Main Results:

• Successful implementation of CRISPR-Cas9 genome editing in yeast.
• Demonstration of both NHEJ and HDR repair mechanisms for precise genetic modifications.
• Completion of the entire genome editing procedure, including verification, within 2-3 weeks.

Conclusions:

• CRISPR-Cas9 technology provides a powerful tool for yeast genome engineering.
• The described method facilitates efficient and accurate genetic modifications for biotechnological purposes.
• This rapid genome editing approach accelerates research and development in yeast systems.