ArticleMolecular therapy. Oncology2025
Highly efficient gene knockout in tumor-infiltrating lymphocytes by adenine base editing.
Article in Molecular therapy. Oncology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed.
- Emerging CRISPR Approaches for Countering Immune Evasion: Insight from Recent Studies.International journal of molecular sciences · 2026Review
- Come together: The BE-TILs-Efficient multiplex base editing of tumor-infiltrating lymphocytes.Molecular therapy. Oncology · 2026Article
- CRISPR-screen informed engineered T cell therapies.Frontiers in immunology · 2026Review
- Advances in organoids for personalized medicine: from technological development to clinical application.Frontiers in cell and developmental biology · 2026Review
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Authors and funding
17 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The disruption of immune checkpoints in T cells is a promising tool for improving the efficacy of tumor-infiltrating lymphocyte (TIL) therapy. While CRISPR-Cas9 genome-editing is efficient, Cas9 nucleases induce double-strand DNA breaks and risks improper translocations, inversions, and chromosomal deletions in engineered T cells. Cas9 nickase (nCas9) used in base-editing cuts only a single strand of DNA, reducing genetic aberrations in modified cells. Here, we established a small-scale, good manufacturing practice-compatible adenine base editing (ABE) procedure for both single and dual knockout of co-inhibitory receptors TIM3 and TIGIT in TILs. ABE-mediated conversion of A·T to G·C pairs in TIM3 and TIGIT specific splice-sites led to high knockout efficiency, with negligible insertion-deletion events post editing. Using melanoma and ovarian TILs, we show that target-specific editing by ABE of TIM3 and TIGIT improved (1) TIL fold-expansion during the rapid expansion protocol without adversely impacting phenotype, (2) cytokine production, and (3) serial killing upon co-culture with autologous patient-derived tumor cells
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Registered trials
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