末端脱氧核酸转移酶介导的功能核酸生物传感器研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2018-0502

摘要/Abstract

摘要： 末端脱氧核酸转移酶（TdT酶）是一种DNA聚合酶,可以催化脱氧核苷酸结合到DNA分子的3'羟基端,并且该反应无需特定的模板。目前,基于末端脱氧核酸转移酶可对模板核酸链的末端进行延伸这一特性,搭载不同的信号输出及扩增方式,构建了一系列的生物传感技术,如电化学生物传感器、荧光生物传感器、表面离子共振生物传感器等。对各类传感器的基本设计原理和应用进行了阐述。根据TdT酶的性质设计的一系列生物传感器具有简单、快速、廉价、灵敏度高、特异性好等优点,实现了对金属离子、病原体、蛋白质等的检测。最后对TdT酶介导的生物传感器目前的研究现状进行了总结并且对TdT酶未来的发展方向进行了展望。

关键词: 末端脱氧核酸转移酶, 3'羟基, 脱氧核苷酸, 末端延伸, 生物传感器

Abstract: Terminal deoxynucleotidyl transferase（TdT enzyme）is a DNA polymerase that catalyzes the binding of deoxynucleotides to the 3’ hydroxyl end of a DNA molecule,and for which a specific template is not demanded. Currently,based on the characteristics of terminal deoxynucleotidyl transferases that can extend the ends of the template nucleic acid strands and carried on with different signal outputs and amplification methods,a series of biosensing technologies were constructed,such as electrochemical biosensors,fluorescent biosensor,and surface ion resonance biosensors. This article describes the basic design principles and applications of various sensors. A series of biosensors can be designed for the detection of metal ions,pathogens,and proteins according to the properties of TdT enzymes,with the advantages of simplicity,rapidity,low cost,high sensitivity,and fine specificity. At the end of this article,the current research status on TdT-mediated biosensors is summarized and its future development direction is prospected.

Key words: terminal deoxynucleotidyl transferase, 3'-OH, deoxynucleotide, end extension, biosensor

张媛, 梁嘉慧, 罗云波, 田晶晶, 田洪涛, 许文涛. 末端脱氧核酸转移酶介导的功能核酸生物传感器研究进展[J]. 生物技术通报, 2019, 35(1): 161-169.

ZHANG Yuan, LIANG Jia-hui, LUO Yun-bo, TIAN Jing-jing, TIAN Hong-tao, XU Wen-tao. Research Progress on Functional Nucleic acid Biosensors Mediated by Terminal Deoxynucleotidyl Transferase[J]. Biotechnology Bulletin, 2019, 35(1): 161-169.

参考文献

[1] Motea EA, Berdis AJ.Terminal deoxynucleotidyl transferase:The story of a misguided DNA polymerase[J]. BBA - Proteins and Proteomics, 2010, 1804(5):1151-1166.
[2] Bollum FJ, Potter VR.Incorporation of thymidine into deoxyribonucleic acid by enzymes from rat tissues[J]. J Biol Chem, 1958, 233(2):478-482.
[3] Chang LM.Development of terminal deoxynucleotidyl transferase activity in embryonic calf thymus gland[J]. Biochemical & Biophysical Research Communications, 1971, 44(1):124-131.
[4] Barr RD, Sarin PS, Perry SM.Terminal transferase in human bone-marrow lymphocytes[J]. Lancet, 1976, 307(7958):508-509.
[5] Srivastav BIS.Association of therminal deoxynucleotidyl transferase activity with chromatin from plant tissue[J]. Biochemical & Biophysical Research Communications, 1972, 48(2):270-273.
[6] Khoury JD, Medeiros LJ, Manning JT, et al.CD56(+)TdT(+)blastic natural killer cell tumor of the skin:a primitive systemic malignancy related to myelomonocytic leukemia[J]. Cancer, 2010, 94(9):2401-2408.
[7] Coleman MS, Hutton JJ, De SP, et al.Terminal deoxyribonucleotidyl transferase in human leukemia[J]. Biochemical & Biophysical Research Communications, 1975, 62(2):367-375.
[8] Gregoire KE, Goldschneider I, Barton RW, et al.Ontogeny of terminal deoxynucleotidyl transferase-positive cells in lymphohemopoietic tissues of rat and mouse[J]. Journal of Immunology, 1979, 123(3):1347-1352.
[9] Bollum FJ.Terminal deoxynucleotidyl transferase as a hematopoietic cell marker[J]. Blood, 1979, 54(6):1203-1215.
[10] Sachs Z, Aasen G, Dolan M, et al.Double- and triple-hit lymphomas can present with features suggestive of immaturity, including TdT expression, and create diagnostic challenges[J]. Leukemia & Lymphoma, 2016, 57(11):2626-2635.
[11] Sur M, Alardati H, Ross C, et al.TdT expression in Merkel cell carcinoma:potential diagnostic pitfall with blastic hematological malignancies and expanded immunohistochemical analysis[J]. Mod Pathol, 2007, 20(11):1113-1120.
[12] Hutton JJ, Coleman MS.Terminal deoxynucleotidyl transferase measurements in the differential diagnosis of adult leukaemias[J]. Br J Haematol, 2010, 34(3):447-456.
[13] Sarin PS, Anderson PN, Gallo RC.Terminal deoxynucleotidyl transferase activities in human blood leukocytes and lymphoblast cell lines:high levels in Lymphoblast cell lines and in blast cells of some patients with chronic myelogenous leukemia in acute phase[J]. Blood, 1976, 47(1):11-20.
[14] Mccaffrey R, Harrison TA, et al.Terminal deoxynucleotidyl transferase activity in human leukemic cells and in normal human thymocytes[J]. N Engl J Med, 1975, 292(15):775-780.
[15] Fowler JD, Suo Z.Biochemical, structural, and physiological characterization of terminal deoxynucleotidyl transferase[J]. Cheminform, 2006, 37(37):2092-2110.
[16] Aravind L, Koonin EV.DNA polymerase β-like nucleotidyltransfe-rase superfamily:Identification of three new families, classification and evolutionary history[J]. Nucleic Acids Research, 1999, 27(7):1609-1618.
[17] Chang LMS, Bollum FJ.Deoxynucleotide polymerizing enzymes of calf thymus gland:IV. Homogeneous terminal deoxynucleotidyl transferase[J]. J Biol Chem, 1971, 246(4):909-916.
[18] Sabbioni E. metalloenzyme nature of calg thymus deoxynucleotidyl transferase[J]. FEBS Lett, 1976, 71(2):233-235.
[19] Srivastava BI.Deoxynucleotide-polymerizing enzyme activities in T- and B-cells of acute lymphoblastic leukemia origin[J]. Cancer Research, 1976, 36(5):1825-1830.
[20] Delarue M, Boulé JB, et al.Crystal structures of a template-indepe-ndent DNA polymerase:murine terminal deoxynucleotidyltransfe-rase[J]. EMBO J, 2002, 21(3):427-439.
[21] Liu Z, Li W, Nie Z, et al.Randomly arrayed G-quadruplexes for label-free and real-time assay of enzyme activity[J]. Chem Commun, 2014, 50(52):6875-6878.
[22] Anne A, Bonnaudat C, Demaille C, et al.Enzymatic redox 3’-end-labeling of DNA oligonucleotide monolayers on gold surfaces using terminal deoxynucleotidyl transferase(TdT)-mediated single base extension[J]. J Am Chem Soc, 2007, 129:2734-2735.
[23] Chi BZ, Liang RP, Zhang L, et al.Sensitive and homogeneous microRNA detection using branched cascade enzymatic amplification[J]. Chem Commun, 2015, 51:10543-10546.
[24] Wu H, Liu S, Jiang J, et al.A sensitive electrochemical biosensor for detection of DNA methyltransferase activity by combining DNA methylation-sensitive cleavage and terminal transferase-mediated extension[J]. Chem Commun, 2012, 48(50):6280-6282.
[25] Wan Y, Wang P, Su Y, et al.Ultrasensitive electrochemical DNA sensor based on the target induced structural switching and surface-initiated enzymatic polymerization[J]. Biosens Bioelectron, 2014, 55(9):231-236.
[26] Ma C, et al.Label-free monitoring of DNA methyltransferase activity based on terminal deoxynucleotidyl transferase using a thioflavin T probe[J]. Mol Cellu Probes, 2016, 30(2):118-121.
[27] Xu F, Luo L, Shi H, et al.label-free and sensitive microRNA detection based on a target recycling amplification-integrated superlong poly(thymine)-hosted copper nanoparticle strategy[J]. Analytica Chimica Acta, 2018, 1010:54-61.
[28] Lu L, Wang M, Liu LJ, et al.A luminescence switch-on probe for terminal deoxynucleotidyl transferase(TdT)activity detection by using an iridium(III)-based i-motif probe[J]. Chem Commun, 2015, 51(49):9953-9956.
[29] Leung KH, He B, Yang C, et al.Development of an aptamer-based sensing platform for metal ions, proteins, and small molecules through terminal deoxynucleotidyl transferase induced G-quadruplex formation[J]. ACS Appl Mater Interfaces, 2015, 7(43):24046-24052.
[30] Wang Y, Chen JT, Yan XP.Fabrication of transferrin functionalized gold nanoclusters/graphene oxide nanocomposite for turn-on near-infrared fluorescent bioimaging of cancer cells and small animals[J]. Anal Chem, 2013, 85(4):2529-2535.
[31] Song C, Yang X, Wang K, et al.label-free and non-enzymatic detection of DNA based on hybridization chain reaction amplification and dsDNA-templated copper nanoparticles[J]. Analytica Chimica Acta, 2014, 827(3):74-79.
[32] Yin J, He X, Wang K, et al.Label-free and turn-on aptamer strategy for cancer cells detection based on a dna-silver nanocluster fluorescence upon recognition-induced hybridization[J]. Anal Chem, 2013, 85(24):12011-12019.
[33] Yuan Y, Li W, Liu Z, et al.A versatile biosensing system for DNA-related enzyme activity assay via the synthesis of silver nanoclusters using enzymatically-generated DNA as template[J]. Biosens Bioelectron, 2014, 61(6):321-327.
[34] Hu Y, Zhang Q, Guo Z, et al.In situ grown DNA nanotail-templated silver nanoclusters enabling label-free electrochemical sensing of terminal deoxynucleotidyl transferase activity[J]. Biosens Bioelectron, 2017, 98:91-99.
[35] Zhou F, Cui X, et al.Fluorometric determination of the activity and inhibition of terminal deoxynucleotidyl transferase via in-situ formation of copper nanoclusters using enzymatically generated DNA as template[J]. Microchim Acta, 2017(3):773-779.
[36] Zhang Q, Hu Y, Xu L, et al.Signal-on electrochemical assay for label-free detection of TdT and BamHI activity based on grown DNA nanowire-templated copper nanoclusters[J]. Analytical & BioAnal Chem, 2017, 409(28):6677-6688.
[37] Hu Y, Shen Q, Li W, et al.A TdT-mediated cascade signal amplification strategy based on dendritic DNA matrix for label-free multifunctional electrochemical biosensing[J]. Biosens Bioelectron, 2015, 63:331-338.
[38] Wei W, Liu S, Sun X, et al.Ultrasensitive electrochemical DNAzyme sensor for lead ion based on cleavage-induced template-independent polymerization and alkaline phosphatase amplification[J]. Biosens Bioelectron, 2016, 83:33-38.
[39] Tian J, et al.Visual single cell detection of dual-pathogens based on multiplex super PCR(MS-PCR)and asymmetric tailing HCR(AT-HCR)[J]. Sens Actuators B Chem, 2018, 260:870-876.
[40] Miao M, et al.Terminal deoxynucleotidyl transferase-induced DNA-
zyme nanowire sensor for colorimetric detection of lipopolysaccha-rides[J]. Sens Actuators B Chem, 2018, 256:790-796.
[41] Zhang Y, Li Q, Li C, et al.Label-free and high-throughput bioluminescent detection of uracil-DNA glycosylase in cancer cells through tricyclic cascade signal amplification[J]. Chem Commun, 2018, 51:6991-6994.
[42] Yuan PX, Deng SY, Zheng CY, et al.In situ formed copper nanoparticles templated by TdT-mediated DNA for enhanced SPR sensor-based DNA assay[J]. Biosens Bioelectron, 2017, 97:1-7.