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Updated: Jul 2, 2025

Selecting and Isolating Colonies of Human Induced Pluripotent Stem Cells Reprogrammed from Adult Fibroblasts

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Ultra-high efficiency T cell reprogramming at multiple loci with SEED-Selection.

Christopher R Chang^1,2,3,4, Vivasvan S Vykunta^1,2,3,4, Daniel B Goodman^1,2,5

¹Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA.

Biorxiv : the Preprint Server for Biology

|February 19, 2024

Summary

This summary is machine-generated.

A novel one-step method, Synthetic Exon/Expression Disruptors (SEEDs), efficiently enriches gene-edited T cells. This process improves purity for complex cellular therapies by removing unwanted cells.

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Area of Science:

Cellular immunotherapy
Gene editing technologies
Biomanufacturing processes

Background:

Current T cell therapies often result in heterogeneous cell mixtures due to incomplete engineering.
Manufacturing challenges limit the clinical application of complex, multiplex gene-edited T cell therapies.

Approach:

Developed Synthetic Exon/Expression Disruptors (SEEDs) for one-step enrichment of gene-edited cells.
SEEDs link transgene integration to endogenous surface protein disruption for immunomagnetic depletion (SEED-Selection).
Designed SEEDs to modify T cell specificity, co-receptor, and MHC expression loci.

Key Points:

Achieved up to 98% purity for single locus modifications and 90% purity for six simultaneous edits (3 knock-ins, 3 knockouts).
SEED-Selection effectively removes non-modified and partially edited cells.
The method is compatible with existing clinical manufacturing workflows.

Conclusions:

SEEDs offer a simple and adaptable approach for producing highly pure, complex gene-edited T cells.
This technology facilitates the development of next-generation cellular therapies.
Facilitates the production of complex gene-edited cell therapies.