title: High-throughput functional microRNAs profiling by recombinant AAV-based microRNA sensor arrays.
abstract: BACKGROUND
  microRNAs (miRNAs) are small and non-coding RNAs which play critical roles in physiological and pathological processes. A number of methods have been established to detect and quantify miRNA expression. However, method for high-throughput miRNA function detection is still lacking.
  
  
  RESULTS
  We describe an adeno-associated virus (AAV) vector-based microRNA (miRNA) sensor (Asensor) array for high-throughput functional miRNA profiling. Each Asensor contains a Gaussia luciferase (Gluc) and a firefly luciferase (Fluc) expression cassette to sense functional miRNA and to serve as an internal control respectively. Using this array, we acquired functional profiles of 115 miRNAs for 12 cell lines and found "functional miRNA signatures" for several specific cell lines. The activities of specific miRNAs including the let-7 family, miR-17-92 cluster, miR-221, and miR-222 in HEK 293 cells were compared with their expression levels determined by quantitative reverse transcriptase polymerase chain reaction (QRT-PCR). We also demonstrate two other practical applications of the array, including a comparison of the miRNA activity between HEK293 and HEK293T cells and the ability to monitor miRNA activity changes in K562 cells treated with 12-O-tetradecanoylphorbol-13-acetate (TPA).
  
  
  CONCLUSIONS
  Our approach has potential applications in the identification of cell types, the characterization of biological and pathological processes, and the evaluation of responses to interventions.
authors.affiliation: College of Life Science, Jilin University, Changchun, Jilin, China.
authors.forename: Wenhong
authors.initials: W
authors.lastname: Tian
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Dong
authors.forename: Xuerong
authors.initials: X
authors.lastname: Liu
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.forename: Zheyue
authors.initials: Z
authors.lastname: Dong
authors.forename: Wei
authors.initials: W
authors.lastname: Shen
authors.forename: Gang
authors.initials: G
authors.lastname: Zheng
authors.forename: Jianxin
authors.initials: J
authors.lastname: Lu
authors.forename: Jinzhong
authors.initials: J
authors.lastname: Chen
authors.forename: Yue
authors.initials: Y
authors.lastname: Wang
authors.forename: Zhijian
authors.initials: Z
authors.lastname: Wu
authors.forename: Xiaobing
authors.initials: X
authors.lastname: Wu
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation*; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D000229:Dependovirus / Q000187:drug effects / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000187:drug effects; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D046228:Microarray Analysis / Q000295:instrumentation*; D011995:Recombination, Genetic / Q000187:drug effects / Q000235:genetics*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D035683:MicroRNAs; D013755:Tetradecanoylphorbol Acetate