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Genetic Screens

Genetic Screens

Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which...

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Updated: Mar 20, 2026

Mating-based Overexpression Library Screening in Yeast

Mating-based Overexpression Library Screening in Yeast

Published on: July 6, 2018

Using Yeast Transposon-Insertion Libraries for Phenotypic Screening and Protein Localization.

¹Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan 48109-1048.

Cold Spring Harbor Protocols

|June 3, 2016

Summary

This study introduces a versatile transposon-insertion library for yeast, enabling efficient phenotypic screening and protein localization studies. This method offers a streamlined alternative to traditional genetic approaches for creating gene disruptions and tagged alleles.

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Last Updated: Mar 20, 2026

Mating-based Overexpression Library Screening in Yeast

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Published on: July 6, 2018

Competitive Genomic Screens of Barcoded Yeast Libraries

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High-throughput Yeast Plasmid Overexpression Screen

High-throughput Yeast Plasmid Overexpression Screen

Published on: July 27, 2011

Area of Science:

Molecular Biology
Yeast Genetics
Functional Genomics

Background:

Traditional methods for creating gene deletions and fluorescent protein fusions in yeast are often labor-intensive.
There is a need for efficient, large-scale mutagenesis tools for functional genomics research.

Purpose of the Study:

To detail a protocol for utilizing a transposon-insertion library for phenotypic screening and protein localization in yeast.
To present a streamlined alternative to labor-intensive targeted approaches for generating mutant alleles.

Main Methods:

Generation of a multipurpose transposon-insertion library in yeast via shuttle mutagenesis.
Introduction of insertion alleles into yeast strains via homologous recombination.
Phenotypic screening of insertion mutants.
Identification of transposon insertion sites using inverse PCR.
Generation of epitope-tagged alleles for protein localization studies using Cre-lox recombination and indirect immunofluorescence.

Main Results:

The transposon-insertion library facilitates efficient gene disruption and the creation of in-frame epitope-tagged alleles.
Phenotypic screening of mutants is readily achievable.
Protein localization can be determined using standard immunofluorescence techniques.
Inverse PCR enables efficient identification of insertion sites.

Conclusions:

Transposon-insertion libraries provide an informative resource for large-scale mutagenesis in yeast.
This approach simplifies the construction of deletion alleles and epitope-tagged proteins.
The protocol offers a powerful tool for functional genomics and understanding protein localization.