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. 2025 Feb 25;27(6):1443–1457. doi: 10.1093/neuonc/noaf055

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© The Author(s) 2025. Published by Oxford University Press on behalf of the Society for Neuro-Oncology.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.

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CRISPRoff repression of MGMT exhibits long-term stability. — CRISPRoff repression of MGMT exhibits long-term stability. (A) RT-qPCR of MGMT mRNA normalized to RPLP0 housekeeping gene, in monoclonal LN18 cells at the indicated number of days following delivery of CRISPRoff and the indicated sgRNA(s). (B) RT-qPCR of MGMT mRNA as in (A) for LN18 cells continuously passaged long term. (C-D) RT-qPCR of MGMT mRNA for T98G monoclonal populations continuously passaged long term following delivery of CRISPRoff and the indicated sgRNA(s). (E) Western blot against MGMT protein in monoclonal LN18 cells at the indicated number of days following delivery of CRISPRoff and the indicated sgRNA. (F) Western blot against MGMT protein as in (E) for LN18 cells delivered with a pool of all 3 sgRNAs against MGMT. (G) Western blot against MGMT protein in monoclonal LN18 cells continuously passaged long term following delivery of CRISPRoff and the indicated sgRNA. (H-I) Western blot against MGMT protein in T98G monoclonal populations continuously passaged long term following delivery of CRISPRoff and the indicated sgRNA(s).