Abstract

Despite current scientific and medical advancements, we yet lack the knowledge necessary to develop targeted therapies for many diseases, like cancer, for which the precise mechanisms of action are unknown due to technological limitations. Current DNA labelling techniques require the killing and fixation of cells, and often have high noise-to-signal ratios, non-specificity and off-target effects. To understand the mechanism of action of diseases accurately, we need a technology applicable to viable observable cells with minimal noise, high specificity and no off-target effects. Through our studies we developed such a visualization approach that uses a modified form of the CRISPR/Cas9 system, conventionally used for genetic editing, and targets DNA and nuclear elements with the afore mentioned precisions. Some clinically applied treatments like chemotherapy are non-specific; they affect both healthy and diseased tissues. The technique we developed allows us to better understand the pathogenesis of disease-causing nuclear changes by labelling, targeting, observing and imaging them. Consequently, we could develop disease-precise therapies without affecting surrounding healthy tissues. While this tool was used on cervical cancer cells, it may be applied to any living cell. We hope that this approach leads to advancements in the field of precision medicine and would help overcome challenges associated with the development of targeted therapies. Importantly, this approach is of comparable low cost and high efficiency, rendering it economically favourable.