Citation: ZHOU Xiao-Yu, ZHAO Jian-Wei, MA Gui-Ming, JIA Hong-Xia. A Cationic Conjugated Polymer-Based Fluorescence Resonance Energy Transfer Strategy for Telomerase Activity Assay Combining with Signal Amplification of Hybridization Chain Reaction[J]. Chinese Journal of Analytical Chemistry, ;2019, 47(7): 1006-1013. doi: 10.19756/j.issn.0253-3820.181330 shu

A Cationic Conjugated Polymer-Based Fluorescence Resonance Energy Transfer Strategy for Telomerase Activity Assay Combining with Signal Amplification of Hybridization Chain Reaction

College of Chemistry and Environment Science, Key Laboratory of Medicinal Chemistry and Molecular Diagnosis of Ministry of Education, Key Laboratory of Analytical Science and Technology of Hebei Province, Hebei University, Baoding 071002, China

Received Date: 18 May 2018
Revised Date: 14 April 2019

Fund Project: This work was supported by the National Natural Science Foundation of China (No.21405032) and the Natural Science Foundation of Hebei Province (No.B2016201052).

Telomerase is usually activated in tumour or cancel cells. The detection of telomerase activity is of great significance for early diagnosis of human humour and cancers as well as screening telomerase-target anticancer therapy. Based on the fluorescence resonance energy transfer (FRET) between cationic conjugated polymer- poly[(9,9-bis(6'-N,N,N-trimethylammonium)hexyl)fluorenylene phenylene (PFP) and SYBR Green I (SG), a simple and fast telomerase activity assay strategy was developed. In the presence of telomerase, the primer was extended, generating a special DNA with many repeat sequences-(GGTTAG)_n. Then, the telomerase elongated products are fixedly connected with magnetic beads through the specific combination of streptavidin (STV) with biotin. A specific DNA probe-(CTAACC)₂ which is matched with two repeat sequences, is used to hybridize with telomerase elongated product. After the telomerase elongated products form double strands DNA (dsDNA), a fluorescent dye SG is added to the hybridization solution and embed into the dsDNA. PFP is added finally. PFP, with positive charges, is a water soluble cationic polymer. It can adsorb on the dsDNA by electrostatic interaction, generating FRET from PFP to SG. So, quantitative determination of telomerase activity can be achieved by detection of FRET efficiency. The telomerase activity in 3×10⁵Hela cells has been obviously detected. To improve the detection sensitivity, hybridization chain reaction (HCR) as signal amplification technology has been used to couple with FRET. The telomerase activity in 6×10⁴ Hela cells has been detected. The detection sensitivity is improved higher almost one order of magnitude.
- Telomerase,
- Activity,
- Cationic conjugated polymer,
- Fluorescence resonance energy transfer,
- Hybridization chain reaction,
- Enzyme-free signal amplification
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