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Cancer therapy with a CRISPR-assisted telomerase-activating gene expression system

Abstract

Cancer is caused by a series of alterations in genome and epigenome and exists in multiple complex forms, making it difficult to be prevented and/or treated. Telomerase, an enzyme responsible for the maintenance of telomere, is silent in most normal somatic cells but activated in 90% of cancer cells, making it an excellent target for cancer therapy. Therefore, various telomerase activity inhibitors have been developed to treat cancer but all failed due to side effects. Here we acted oppositely to develop a cancer gene therapy named telomerase-activating gene expression (Tage) system by utilizing the telomerase activity in cancer cells. The Tage system consisted of an effector gene expression vector that carried a 3ʹ telomerase-recognizable stick end and an artificial transcription factor expression vector that could express dCas9-VP64 and an sgRNA targeting telomere repeat sequences. By using Cas9 as an effector gene, the Tage system effectively killed various cancer cells, including HepG2, HeLa, PANC-1, MDA-MB-453, A549, HT-29, SKOV-3, Hepa1-6, and RAW264.7, without affecting normal cells MRC-5, HL7702, and bone marrow mesenchymal stem cell (BMSC). More importantly, a four-base 3ʹ stick end produced by the homothallic switching endonuclease in cells could be recognized by telomerase, allowing the Tage system to effectively kill cancer cells in vivo. The Tage system could effectively and safely realize its in vivo application by using adeno-associated virus (AAV) as gene vector. The virus-loaded Tage system could significantly and specifically kill cancer cells in mice by intravenous drug administration without side effects or toxicity.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (61571119) and the National Key Research and Development Program of China (2017YFA0205502).

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JWang conceived the study and designed the experiments. WD designed and performed main experiments. XX, DW and JWu prepared reagents and performed partial experiments. JWang and WD wrote the manuscript with support from all authors.

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Correspondence to Jinke Wang.

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Dai, W., Xu, X., Wang, D. et al. Cancer therapy with a CRISPR-assisted telomerase-activating gene expression system. Oncogene 38, 4110–4124 (2019). https://doi.org/10.1038/s41388-019-0707-8

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  • DOI: https://doi.org/10.1038/s41388-019-0707-8

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