The discovery of the bacterial Clustered, Regularly Interspaced Short Palindromic Repeats (CRISPR) and associated Cas9 protein, and its ability to edit genes in mammalian cells is poised to revolutionise our ability to treat genetic diseases, particularly in the cancer setting where the driver genes are known. Indeed, CRISPR/Cas9 was shown to effectively edit any gene of interest with high efficiency and at low cost. However, a major challenge to treating cancer is the heterogeneity of the

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... ic alterations and the ongoing accumulation of new mutations that enable cancers to survive and drug resistance to emerge. With the increasing knowledge of cancer biology, thanks to the advances in DNA sequencing technologies, the low cost, and the ability to edit genes with CRISPR/Cas9 with high efficiency, it is now possible to develop more targeted anticancer therapeutics with promising outcomes. Despite the exceptional CRISPR/Cas9 gene-editing features, the early studies found several limitations to its potential use as a human gene therapy, such as the targeting specificity of CRISPR/Cas9, the in vivo delivery of the treatment with minimal systemic toxicity, the in vivo efficacy under immunocompetent conditions, the immunogenicity of CRISPR/Cas9 and the delivery vehicle when delivered systemically, and whether the introduced edits would induce an immune response. Particularly for cancer treatment, challenges can be broadly categorized into: the in vivo efficacy of CRISPR/Cas9 therapeutics, the in vivo delivery of the treatment to target tissue with high transfection efficiency, and the heterogeneity of genetic mutations in cancer, and thus targeting a known gene may not be enough to cure cancer. To enable the utilisation of CRISPR/Cas9 as an anticancer therapy, the above-mentioned challenges need to be addressed. We utilised the well-established Human Papillomavirus (HPV)-driven cervical cancer models due to their addiction on the expression of HPV oncogenes, namely E6 and E7, for their survival and progression, and t [...]