Journal
TALANTA
Volume 161, Issue -, Pages 804-811Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.talanta.2016.09.040
Keywords
Integrated microfluidics; Single-cell multiplex qPCR; Genotoxicity assay
Categories
Funding
- Youth Foundation of Fuzhou University [510215, 510216]
- National Natural Science Foundation of China [61604042]
- National Overseas Scholarship from the China Scholarship Council [201206120110]
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With the development of large-scale biologic databases, precision medicine is becoming a frontier in biomedical research. As a main focus of precision medicine study, cancer has been widely accepted as a disease born out of inherited genetic variations or accumulating genomic damage. At the single-cell level, microfluidics or lab-on-a chip technology for cancer study is an emerging tool for improving risk assessment, diagnostic categories and therapeutic strategies. This work presents a multi-layer microchip for single-cell gene expression profiling. Treated by three drug reagents (i.e. methyl methanesulfonate, docetaxel and colchicine) with varied concentrations and time lengths, individual human breast cancer cells (MCF-7) are then lysed on-chip, and the released mRNA templates are captured and reversely transcribed into cDNA on microbead surface. Three genes (GAPDH, CDKN1A, AURKA) are amplified and quantified simultaneously through triplex real-time polymerase chain reactions (qPCR). Readout per run is set to be eighteen, and can be further improved following same approach. The microchip is able to integrate all steps of single-cellgene expression profiling, and provide precision study of drug induced genotoxicity with reduced reagents consumption per reaction and instrumental cost.
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