A user-friendly and streamlined protocol for CRISPR/Cas9 genome editing in budding yeast.

Area of Science:

• Molecular Biology
• Genetics
• Biotechnology

Background:

• CRISPR/Cas9 is a powerful tool for precise, marker-less genome editing.
• Efficient and accessible genome editing protocols are crucial for advancing yeast research.

Purpose of the Study:

• To provide a detailed, robust, and streamlined protocol for CRISPR/Cas9 genome editing in Saccharomyces cerevisiae.
• To make CRISPR/Cas9 technology more accessible to yeast research laboratories.

Main Methods:

• Utilized the MoClo-Yeast Toolkit for CRISPR/Cas9 system assembly.
• Developed a step-by-step protocol from single-guide RNA (sgRNA) design to verification.
• Provided preassembled plasmids for sgRNA cloning.

Main Results:

• Successfully established a detailed and streamlined CRISPR/Cas9 genome editing protocol for yeast.
• The protocol simplifies the process from sgRNA design to validating genome editing events.
• Preassembled plasmids facilitate easier sgRNA cloning and integration into the workflow.

Conclusions:

• The presented protocol enhances the accessibility and efficiency of CRISPR/Cas9 genome editing in Saccharomyces cerevisiae.
• This resource empowers yeast research groups to perform accurate, marker-less genome modifications.
• The MoClo-Yeast Toolkit-based approach streamlines genetic engineering in yeast.