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ORIGINAL RESEARCH article

Front. Bioeng. Biotechnol.
Sec. Cell and Gene Therapy
Volume 12 - 2024 | doi: 10.3389/fbioe.2024.1417070
This article is part of the Research Topic CRISPR tailored gene editing – the real promise to treat human disease View all 5 articles

Assessing a single-cell multi-omic analytic platform to characterize ex vivo-engineered T cell therapy products

Provisionally accepted
Maryam Moshref Maryam Moshref 1*Jerry Lo Jerry Lo 1Andrew Mckay Andrew Mckay 1Julien Camperi Julien Camperi 1Joseph Schroer Joseph Schroer 1Norikiyo Ueno Norikiyo Ueno 2Shu Wang Shu Wang 2Saurabh Gulati Saurabh Gulati 2Somayeh Tarighat Somayeh Tarighat 1Steffen Durinck Steffen Durinck 1Ho-Young Lee Ho-Young Lee 1Dayue Chen Dayue Chen 1*
  • 1 Genentech Inc., San Francisco, United States
  • 2 Mission Bio, South San Francisco, United States

The final, formatted version of the article will be published soon.

    Genetically engineered CD8 + T cells are being explored for the treatment of various cancers. Analytical characterization represents a major challenge in the development of genetically engineered cell therapies, especially assessing the potential off-target editing and product heterogeneity. As conventional sequencing techniques only provide information at the bulk level, they are unable to detect off-target CRISPR translocation or editing events occurring in minor cell subpopulations. Here we report the analytical development of a single-cell multi-omic DNA and Protein assay to characterize genetically engineered cell products for safety and genotoxicity assessment. We were able to quantify on-target edits, off-target events, and potential translocations at the targeting loci with per-cell granularity, providing important characterization data of the final cell product.Conclusion: Single cell multi-omics approach provides the resolution required to understand the composition of cellular products and identify critical quality attributes (CQAs).

    Keywords: single-cell1, multi-omic2, engineered CD8 + T cells3, CRISPR4, heterogeneity5, final cell product6

    Received: 13 Apr 2024; Accepted: 11 Jul 2024.

    Copyright: © 2024 Moshref, Lo, Mckay, Camperi, Schroer, Ueno, Wang, Gulati, Tarighat, Durinck, Lee and Chen. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence:
    Maryam Moshref, Genentech Inc., San Francisco, United States
    Dayue Chen, Genentech Inc., San Francisco, United States

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.