RNAi-Enhanced Logic Circuit for Cancer Specific Detection and Destruction.

摘要

Modern breast cancer therapies utilize non-specific approaches to kill or remove cancerous cells, inflicting significant collateral damage to healthy cells. In response to the need for highly targeted detection and destruction of cancerous cells, we propose to implement multi-input genetic circuits that act as cell state classifiers based on mRNA or microRNA expression profiling. The mRNA sensing project is focused on the MCF-7 breast adenocarcinoma cell line. MCF-7 cells overexpress Gata3, NPY1R and TFF1 mRNA relative to healthy cells. Based on our bioinformatics analysis, taking into account the three biomarkers allows for dramatically improved specificity in comparison to targeting single genes. We therefore design a three-input AND gate that triggers a response only when all three biomarkers are expressed above a defined threshold. In second approach we implement transcriptional/post- transcriptional regulatory circuit that senses expression levels of a customizable set of endogenous microRNAs and computes whether to trigger a response if the expression levels match a pre-determined profile of interest. We have created a circuit that computes a complex abstract logic (miR1 AND miR2+3 AND NOT miR4 AND NOTmiR5 AND NOT miR6) and selectively triggers output response in HeLa but not in other cells.