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Updated: Jun 22, 2025
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Production of Human CRISPR-Engineered CAR-T Cells
Published on: March 15, 2021
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An aptamer-mediated base editing platform for simultaneous knockin and multiple gene knockout for allogeneic CAR-T
Immacolata Porreca1, Robert Blassberg1, Jennifer Harbottle1
1Revvity, 8100 Cambridge Research Park, Cambridge CB25 9TL, UK.
Summary
A new Pin-point base editing platform precisely modifies genes in human T cells for advanced therapies. This method reduces unwanted DNA damage compared to CRISPR-Cas9, enabling safer cell therapies.
Area of Science:
- Biotechnology
- Molecular Biology
- Genetics
Background:
- Gene editing advances adoptive cellular therapies.
- Conventional CRISPR-Cas9 causes DNA double-strand breaks (DSBs), leading to genotoxicity and unwanted genomic alterations.
Purpose of the Study:
- To evaluate a novel RNA aptamer-mediated Pin-point base editing platform for complex genome editing in human primary T cells.
- To compare the safety and efficiency of the Pin-point platform against conventional CRISPR-Cas9.
Main Methods:
- Application of a modular RNA aptamer-mediated Pin-point base editing platform.
- Simultaneous introduction of multiple gene knockouts and site-specific transgene integration.
- Editing in human primary T cells.
Main Results:
- High editing efficiency and purity were achieved at all target sites.
- Significantly reduced frequency of chromosomal translocations compared to CRISPR-Cas9.
- Successful site-specific knockin of a chimeric antigen receptor and multiplex gene knockout in a single intervention.
Conclusions:
- The Pin-point platform enables efficient and precise complex genome editing.
- This technology offers a safer alternative to CRISPR-Cas9 by minimizing genotoxicity.
- The Pin-point platform has significant potential for various advanced cell therapies.
