title: Colorimetric detection of gene transcript by target-induced three-way junction formation.
abstract: Gene transcript often varies by alternative splicing, which plays different biological role that results in diversity of gene expression. Therefore, a simple and accurate identification of targeted transcript variant is of prime importance to achieve a precise molecular diagnosis. In this work, we presented a three-way junction based system where two split G-quadruplex forming sequences were coupled into two probes. Only upon the introduction of target gene transcript that offering a specific recognizable splicing site did the two probes assembled into three way junction conformation in a devised process, thus providing a functional G-quadruplex conformation that greatly enhanced hemin peroxidation. A notable resolution for gene splicing site detection was achieved. The detection limitation by colorimetric assay was 0.063μM, and this system has been proved to discriminate even in a single base false level around splicing site (about 3 times of single mismatched analyte to gain an equal signal by perfect analyte ). Furthermore, recoveries of 78.1%, 88.1%, 104.6% were obtained with 0.75μM, 0.25μM, 0.083μM of target, respectively, showing a capacity to further exploit a simple equipped device for gene transcript detection.
authors.affiliation: Department of Laboratory Medicine, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
authors.forename: Xuchu
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Laboratory Medicine, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Liu
authors.affiliation: Department of Laboratory Medicine, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
authors.forename: Binbin
authors.initials: B
authors.lastname: Yin
authors.affiliation: Department of Laboratory Medicine, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
authors.forename: Pan
authors.initials: P
authors.lastname: Yu
authors.affiliation: Department of Laboratory Medicine, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
authors.forename: Xiuzhi
authors.initials: X
authors.lastname: Duan
authors.affiliation: Department of Blood Transfusion, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
authors.forename: Zhaoping
authors.initials: Z
authors.lastname: Liao
authors.affiliation: Department of Blood Transfusion, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
authors.forename: Chunhua
authors.initials: C
authors.lastname: Liu
authors.affiliation: Department of Laboratory Medicine, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
authors.forename: Yiwen
authors.initials: Y
authors.lastname: Sang
authors.affiliation: Department of Laboratory Medicine, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
authors.forename: Gong
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Department of Laboratory Medicine, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Laboratory Medicine, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China. Electronic address: zrtzh@zju.edu.cn.
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Tao
mesh_terms: D017398:Alternative Splicing; D000951:Antigens, Neoplasm / Q000235:genetics*; D003124:Colorimetry; D018076:DNA, Complementary; D054856:G-Quadruplexes; D006801:Humans; D014158:Transcription, Genetic
chemical_list: D000951:Antigens, Neoplasm; D018076:DNA, Complementary; C479341:prostate cancer antigen 3, human
keywords: Colorimetric detection; Gene transcript; Three way junction formation