Generation and Applications of a DNA Aptamer against Gremlin-1
1
Department of Rheumatology, the First Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061, China
2
Research Center of Reproductive Medicine, Medical School of Xi’an Jiaotong University, Xi’an 710061, China
3
Department of Infectious Diseases, the Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an 710061, China
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Academic Editor: Harri Lönnberg
Molecules 2017, 22(5), 706; https://doi.org/10.3390/molecules22050706
Received: 16 March 2017
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Revised: 15 April 2017
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Accepted: 22 April 2017
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Published: 28 April 2017
(This article belongs to the Special Issue Synthesis and Applications of Oligonucleotide Conjugates)
Gremlin-1, a highly conserved glycosylated and phosphorylated secretory protein, plays important roles in diverse biological processes including early embryonic development, fibrosis, tumorigenesis, and renal pathophysiology. Aptamers, which are RNA or DNA single-stranded oligonucleotides capable of binding specifically to different targets ranging from small organics to whole cells, have potential applications in targeted imaging, diagnosis and therapy. In this study, we obtained a DNA aptamer against Gremlin-1 (G-ap49) using in vitro Systematic Evolution of Ligands by Exponential Enrichment (SELEX). Binding assay and dot-blot showed that G-ap49 had high affinity for Gremlin-1. Further experiments indicated that G-ap49 was quite stable in a cell culture system and could be used in South-Western blot analysis, enzyme-linked aptamer sorbent assay (ELASA), and aptamer-based cytochemistry and histochemistry staining to detect Gremlin-1. Moreover, our study demonstrated that G-ap49 is capable of revealing the subcellular localization of Gremlin-1. These data indicate that G-ap49 can be used as an alternative to antibodies in detecting Gremlin-1.
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Keywords:
aptamer; Gremlin-1; South-Western blot analysis; enzyme-linked aptamer sorbent assay; immunohisto-cytochemical staining
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MDPI and ACS Style
Li, Q.; Huo, Y.; Guo, Y.; Zheng, X.; Sun, W.; Hao, Z. Generation and Applications of a DNA Aptamer against Gremlin-1. Molecules 2017, 22, 706. https://doi.org/10.3390/molecules22050706
AMA Style
Li Q, Huo Y, Guo Y, Zheng X, Sun W, Hao Z. Generation and Applications of a DNA Aptamer against Gremlin-1. Molecules. 2017; 22(5):706. https://doi.org/10.3390/molecules22050706
Chicago/Turabian StyleLi, Qian, Yongwei Huo, Yonghong Guo, Xiaoyan Zheng, Wengang Sun, and Zhiming Hao. 2017. "Generation and Applications of a DNA Aptamer against Gremlin-1" Molecules 22, no. 5: 706. https://doi.org/10.3390/molecules22050706
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