A reversible gene trap collection empowers haploid genetics in human cells

Area of Science:

• Genomics
• Cell Biology
• Human Genetics

Background:

• Knockout collections are essential for studying model organisms like yeast.
• Large-scale knockout collections for human cells are currently lacking.
• Gene-trap mutagenesis offers a method for generating gene knockouts.

Purpose of the Study:

• To establish a platform for generating and isolating gene-trapped human cells.
• To create a comprehensive library of human cell lines with single gene-trap insertions.
• To enable systematic screens for diverse cellular phenotypes in human cells.

Main Methods:

• Utilized gene-trap mutagenesis in near-haploid human cells.
• Developed a platform for generating and isolating individual gene-trapped cells.
• Created a DNA-barcoded library covering 3,396 genes (one-third of the expressed genome).

Main Results:

• Successfully generated a collection of human cell lines with single gene-trap insertions.
• The library includes 3,396 genes and is DNA-barcoded for systematic screening.
• Demonstrated the utility of the collection by examining cellular responses to TNF-α, TGF-β, IFN-γ, and TRAIL.

Conclusions:

• The developed platform provides a valuable resource for studying human cell biology.
• This gene-trapped cell collection enables large-scale functional genomic screens.
• The library facilitates the investigation of cellular responses to various signaling pathways.