Generation and Applications of a DNA Aptamer against Gremlin-1

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...

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Bibliographic Details
Published in:Molecules (Basel, Switzerland) Switzerland), 2017-04, Vol.22 (5), p.706
Main Authors: Li, Qian, Huo, Yongwei, Guo, Yonghong, Zheng, Xiaoyan, Sun, Wengang, Hao, Zhiming
Format: Article
Language:English
Subjects:
Affinity
Animals
Aptamers
Aptamers, Nucleotide - chemical synthesis
Aptamers, Nucleotide - metabolism
Assaying
Base Sequence
Binding
Biological activity
Biological evolution
Cell culture
Cytochemistry
Deoxyribonucleic acid
Diagnosis
DNA
Embryogenesis
Embryonic growth stage
Enrichment
Evolution
Fibrosis
Gremlin protein
HEK293 Cells
Histochemistry
Humans
Imaging
Immunoblotting
In vitro methods and tests
Intercellular Signaling Peptides and Proteins - metabolism
Localization
Mice
Microscopy, Fluorescence
SELEX Aptamer Technique
Single-stranded DNA
Staining
Staining and Labeling
Tumorigenesis
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Summary: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.
ISSN:1420-3049
1420-3049
DOI:10.3390/molecules22050706