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Using Functional Genetics in Haploid Cells for Drug Target Identification.

Jennifer C Volz¹, Nicole Schuller¹, Ulrich Elling²

¹IMBA - Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Vienna, Austria.

Methods in Molecular Biology (Clifton, N.J.)

|March 27, 2019

Summary

Haploid gene trap screens offer a powerful method for drug discovery and identifying genetic interactions. This protocol details a step-by-step approach for performing these screens in mammalian cells, providing an alternative to other screening techniques.

Keywords:

Functional genomics Gene trap Genetic screen Haploid Stem cell

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Area of Science:

Genetics
Drug Discovery
Molecular Biology

Background:

Pooled genetic screens are vital for identifying drug targets and understanding chemogenetic interactions.
Methods like mRNA knockdown, genome editing, and random mutagenesis generate complex cell populations for screening.
Haploid mammalian cell lines enhance the power of random mutagenesis for genetic screening.

Purpose of the Study:

To provide a detailed protocol for conducting haploid gene trap screens.
To present random mutagenesis in haploid cells as a viable alternative to other screening methods.
To facilitate the identification of novel drug targets and chemogenetic interactions.

Main Methods:

Utilizing haploid mammalian cell lines for genetic screening.
Implementing random genome mutagenesis techniques.
Performing a step-by-step gene trap screening protocol.

Main Results:

The protocol enables the generation of highly complex cell populations for genetic screens.
Haploid gene trap screening proves effective for target identification.
This method serves as a practical alternative to CRISPR-based screening.

Conclusions:

Haploid gene trap screens are a powerful and accessible tool for genetic research.
The protocol facilitates drug target identification and the study of genetic interactions.
Random mutagenesis in haploid cells offers a robust approach for screening applications.