Evidence map›Paper›PMID 39998382›Full record

ArticleNeuro-oncology2025

Multiplexed epigenetic memory editing using CRISPRoff sensitizes glioblastoma to chemotherapy.

Katie Lin, Christopher Zou, Akane Hubbard, Sasha Sengelmann, Laine Goudy, I-Ching Wang, Rohit Sharma, Joanna Pak, Kyla Foster, Tomoko Ozawa and 9 more

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Neuro-oncology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.

0numbers the graph read from it
0cells of the map it votes in
16citing 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

16 citing papers in PubMed.

  1. Review
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  5. Advances and clinical potential of epigenome editing.Cellular and molecular life sciences : CMLS · 2026
    Review
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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

19 authors.

Katie LinDepartment of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.
Christopher ZouDepartment of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.
Akane HubbardDepartment of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.
Sasha SengelmannDepartment of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.
Laine GoudyGladstone Institutes-UCSF, Institute of Genomic Immunology, San Francisco, California, USA.
I-Ching WangDepartment of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.
Rohit SharmaDepartment of Bioengineering, University of California Berkeley, Berkeley, California, USA.
Joanna PakDepartment of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.
Kyla FosterDepartment of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.
Tomoko OzawaDepartment of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.
John F de GrootDepartment of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.
Joanna PhillipsDepartment of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.
Harish N VasudevanDepartment of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.
David R RaleighDepartment of Pathology, University of California San Francisco, San Francisco, California, USA.
Alexander MarsonGladstone Institutes-UCSF, Institute of Genomic Immunology, San Francisco, California, USA.
Niren MurthyDepartment of Bioengineering, University of California Berkeley, Berkeley, California, USA.
Luke A GilbertArc Institute, Palo Alto, California, USA.
Mitchel S BergerDepartment of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.
S John LiuDepartment of Neurological Surgery, University of California San Francisco, San Francisco, California, USA.ORCID 0000-0003-1042-0191

Funding

Tissue CoreP50CA097257 · NCI · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI Joseph F Costello · 2002 to 2026
$57.4M
Training Program in Translational Brain Tumor ResearchT32CA151022 · NCI · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI Joseph F Costello · 2010 to 2026
$6.6M
CRISPRNCI NIH HHS P50 CA097257NCI NIH HHS T32 CA151022NIH HHS P50 CA097257
6 · The paper itself

Abstract

backgroundGlioblastoma (GBM) carries a poor prognosis, and new therapeutic strategies are necessary to improve outcomes for patients with this disease. Alkylating chemotherapies including temozolomide (TMZ) and lomustine (CCNU) are critical for treating GBM, but resistance mechanisms, including hypomethylation of O6-methylguanine-DNA methyltransferase (MGMT) promoter, undermine treatment. CRISPRoff is a programmable epigenetic memory editor that can induce stable and heritable gene silencing after transient delivery, and we hypothesize that CRISPRoff could potentiate the activity of TMZ and CCNU through long-term suppression of target genes.

methodsWe transiently delivered CRISPRoff mRNA along with sgRNAs against target genes using both electroporation and lipid nanoparticles (LNPs) into established GBM cell lines, patient-derived primary GBM cultures, and orthotopic GBM xenografts. Gene repression, specificity, and stability were measured by RT-qPCR, Western blot, bisulfite sequencing, and RNA sequencing. Sensitivity to chemotherapies was measured by cell viability dose-response, microscopy, and bioluminescence imaging. Genome-wide mapping of CCNU sensitizers was performed using CRISPRi screens.

resultsCRISPRoff induced complete suppression of MGMT and sensitization to TMZ that was stable for over 8 months of continuous cell propagation. GBM orthotopic tumors treated with CRISPRoff against MGMT demonstrated sensitivity to TMZ in vivo, and CRISPRoff delivery resulted in chemosensitivity in patient-derived primary GBM. Genome-wide CRISPRi screens identified combinatorial genetic vulnerabilities (BRIP1, FANCE) that were targetable by multiplexed CRISPRoff to achieve sensitization to CCNU.

conclusionTransient delivery of a site-specific epigenetic memory can induce stable, complete, and multiplexed suppression of target genes for therapeutic application in GBM.

Indexed as

Brain NeoplasmsCRISPR-Cas SystemsDrug Resistance, NeoplasmEpigenesis, GeneticGene EditingGlioblastomaAnimalsDNA Modification MethylasesDNA Repair EnzymesEpigenetic MemoryHumansLomustineMiceMice, NudeTemozolomideTumor Cells, CulturedDNA Modification MethylasesDNA Repair EnzymesLomustineMGMT protein, humanTemozolomideTumor Suppressor ProteinsCCNUCRISPRepigenetic editingglioblastomatemozolomide

Identifiers

PMID39998382
PMCPMC12309708

What OpenQuestion holds

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LicenceCC BY-NC
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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.