High-throughput single-cell assessment of multiplex gene editing

Multiplex gene editing is becoming more common because base and prime editing don’t depend on double-strand breaks, which makes them more flexible and safer. However, there are still no good assays for accurately measuring multiplex editing. In this webinar, Stefan Radtke, PhD, of Fred Hutch Cancer Center discusses new high-throughput single-cell readouts for tracking gene editing outcomes after autologous hematopoietic stem cell gene therapy.

He describes a single-cell sequencing strategy that reliably captures all possible genotypes across different tissues and blood cell types, at a resolution that bulk sequencing can’t reach.

Topics covered:

• Why bulk sequencing approaches cannot accurately capture gene editing outcomes, and underestimate the frequency of edited cells after transplantation
• Why monitoring multiplex gene editing after gene therapy needs new readouts built on high-throughput single-cell genomics
• How cell-by-cell sequencing captures mono- and bi-allelic editing outcomes, reveals sgRNA-specific editing biases, and can be adapted for other multiplexing strategies, clone tracking and off-target monitoring