Evidence map›Paper›PMID 41376778›Full record

ArticleMolecular therapy. Methods & clinical development2025

CRISPR/methotrexate-integrated strategy for TCR-T cell engineering with reduced chromosome 14 loss.

Jian Xu, Lianghua Shen, Ziyu Chen, Changmeng Zhang, Xiaodan Ding, Qiaomei He, Xiao Zhou, Peiyao Jiang, Jiayu Liu, Lu Li and 9 more

Erratum issuedAbstract read
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In one paragraph

Article in Molecular therapy. Methods & clinical development, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Not yet cited in PubMed.

0numbers the graph read from it
0cells of the map it votes in
0citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

19 authors.

Jian XuDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Lianghua ShenDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Ziyu ChenDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Changmeng ZhangDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Xiaodan DingDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Qiaomei HeDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Xiao ZhouDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Peiyao JiangDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Jiayu LiuDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Lu LiDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Jinan FangDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Fanlin LiDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Liping WanDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Xueying DingEngineering Technology Research Center of Cell Therapy and Clinical Translation, Science and Technology Committee of Shanghai Municipality, Shanghai 201620, China.
Yuqin YangAnimal Core Facility, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, China.
Xiaorui WangDepartment of Laboratory Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, China.
Pengran WangDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Xianmin SongDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.
Yan ZhangDepartment of Hematology, Shanghai General Hospital, Shanghai JiaoTong University School of Medicine, Shanghai 200080, China.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Adoptive T cell therapy, particularly T cell receptor-engineered T (TCR-T) cell therapy, holds promise for cancer treatment in solid tumors and hematological malignancies. Conventional lentiviral TCR-T cell therapies face insertional mutagenesis risks, while CRISPR-mediated T cell receptor α constant (TRAC) locus targeting suffers from suboptimal knockin efficiency and chromosome 14 loss. To address these challenges, we introduced a non-viral strategy combining CRISPR-Cas9 electroporation with methotrexate (MTX) metabolic selection. Primary human T cells were engineered to integrate a CMV-pp65-specific TCR and an MTX-resistant dihydrofolate reductase (DHFR)-FS cassette into the TRAC locus via homology-directed repair (HDR). Systematic optimization of electroporation timing, buffer systems, and HDR enhancers achieved initial TCR integration efficiency of ∼20%. Subsequent 6-day MTX treatment enriched engineered cells to ∼70% purity while selectively depleting unedited and chromosomally aberrant clones. Fluorescence

Indexed as

adoptive cell therapyCRISPRelectroporationimmunotherapyT cellT cell receptor

Identifiers

PMID41376778
PMCPMC12686916

What OpenQuestion holds

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.