Efficient Precision Genome Editing in iPSCs via Genetic Co-targeting with Selection

Area of Science:

• Stem cell biology
• Genome engineering
• Molecular genetics

Background:

• Identifying homology-directed repair (HDR) modified cells is a major bottleneck in genome editing of induced pluripotent stem cells (iPSCs).
• Current methods for isolating HDR-edited cells are often laborious and inefficient, hindering the development of disease models.

Purpose of the Study:

• To develop a more efficient method for isolating homology-directed repair (HDR)-modified cells in induced pluripotent stem cells (iPSCs).
• To accelerate the generation of iPSC-based disease models by improving the selection of edited cells.

Main Methods:

• Developed a novel strategy termed "co-targeting with selection" for genome editing in iPSCs.
• This method leverages the isolation of cells with a selectable, HDR-mediated editing event at one genomic locus to enrich for HDR-mediated edits at additional loci.

Main Results:

• The co-targeting with selection strategy significantly improves the probability of isolating iPSCs bearing desired HDR-mediated variants.
• This approach streamlines the process of generating genetically modified cell lines.

Conclusions:

• Co-targeting with selection offers a more efficient and accelerated approach to isolating HDR-modified iPSCs.
• This method facilitates the rapid production of iPSC-based disease models for research and therapeutic development.