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MX Cassettes for Knocking Out Genes in Yeast.
1Department of Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, North Carolina 27710 john.mccusker@duke.edu.
Cold Spring Harbor Protocols
|April 5, 2017
Summary
This study details MX cassettes for precise Saccharomyces cerevisiae genome editing. These knockout cassettes facilitate gene deletion with various selection markers, simplifying genetic modifications.
Area of Science:
- Molecular Biology
- Yeast Genetics
Background:
- Precise genome modifications in Saccharomyces cerevisiae commonly employ knockout (KO) marker cassettes for gene deletion.
- A variety of KO marker cassettes exist, differing in strain genotype requirements and selection strategies.
Purpose of the Study:
- To provide a focused overview of the widely used MX cassette family for Saccharomyces cerevisiae genome editing.
- To describe the different types of MX cassettes and their associated selection methods, including prototrophy, cytosine or acetamide utilization, and drug resistance.
Main Methods:
- Review and description of various MX cassette types and their selection mechanisms.
- Discussion of cassette recycling ('pop out') and counterselection strategies.
- Consideration of strain genotype and media requirements for cassette application.
Main Results:
- Detailed explanation of MX cassettes, highlighting selections for prototrophy, cytosine/acetamide utilization, and resistance to six drugs.
- Elucidation of strain genotype requirements for specific cassettes.
- Explanation of cassette recycling and counterselection techniques.
Conclusions:
- MX cassettes offer versatile and efficient tools for targeted gene deletion in Saccharomyces cerevisiae.
- Understanding the different MX cassette types and their selection markers is crucial for successful genome engineering.
- The described methods for cassette recycling and counterselection enhance the utility of MX cassettes in complex genetic studies.