title: A Microchip for Integrated Single-Cell Gene Expression Profiling and Genotoxicity Detection.
abstract: Microfluidics-based single-cell study is an emerging approach in personalized treatment or precision medicine studies. Single-cell gene expression holds a potential to provide treatment selections with maximized efficacy to help cancer patients based on a genetic understanding of their disease. This work presents a multi-layer microchip for single-cell multiplexed gene expression profiling and genotoxicity detection. Treated by three drug reagents (i.e., methyl methanesulfonate, docetaxel and colchicine) with varied concentrations and time lengths, individual human cancer cells (MDA-MB-231) are lysed on-chip, and the released mRNA templates are captured and reversely transcribed into single strand DNA. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cyclin-dependent kinase inhibitor 1A (CDKN1A), and aurora kinase A (AURKA) genes from single cells are amplified and real-time quantified through multiplex polymerase chain reaction. The microchip is capable of integrating all steps of single-cell multiplexed gene expression profiling, and providing precision detection of drug induced genotoxic stress. Throughput has been set to be 18, and can be further increased following the same approach. Numerical simulation of on-chip single cell trapping and heat transfer has been employed to evaluate the chip design and operation.
authors.affiliation: School of Mechanical Engineering and Automation, Fuzhou University, Fujian 350116, China. hdong@fzu.edu.cn.
authors.forename: Hui
authors.initials: H
authors.lastname: Dong
authors.affiliation: School of Mechanical Engineering and Automation, Fuzhou University, Fujian 350116, China. sh@fzu.edu.cn.
authors.forename: Hao
authors.initials: H
authors.lastname: Sun
mesh_terms: D002138:Calibration; D045744:Cell Line, Tumor; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D044085:Microfluidics / Q000379:methods*; D012313:RNA / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D059010:Single-Cell Analysis / Q000379:methods*; D013696:Temperature
chemical_list: D012313:RNA
keywords: integrated multiplex RT-qPCR; microfluidics; single-cell analysis