A NOVEL SSDNA APTAMER TARGETING CARCINOEMBRYONIC ANTIGEN: SELECTION AND CHARACTERIZATION
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Date
2022-10-20
Authors
Yunussova, Nigara
Sypabekova, Marzhan
Zhumabekova, Zhazira
Matkarimov, Bakhyt
Kanayeva, Damira
Journal Title
Journal ISSN
Volume Title
Publisher
Biology
Abstract
One of the major causes of a drastically shorter life expectancy and one of the most prevalent
diseases in the world today is cancer. Given the data on the rise in cancer cases throughout the
world, it is obvious that, despite the diagnostic techniques currently being used, there is a pressing
need to develop precise and sensitive techniques for early diagnosis of the disease. A high degree of
affinity and specificity towards particular targets is maintained by the short nucleic acid molecules
known as aptamers. Aptamers outperform antibodies due to their unique benefits, such as their
simplicity in synthesis and modification, lack of toxicity, and long-term stability. Utilizing an accurate
recognition element and a robust signal transduction mechanism, molecular diagnostics can be
extremely sensitive and specific. In this study, development of new single-stranded DNA aptamers
against CEA for use in cancer diagnostics was accomplished using SELEX and NGS methods. As a
result of 12 iterative SELEX rounds, nine aptamer candidates against CEA were developed. NGS
comparative analysis revealed that round twelve had an enriched number of aptamers that were
specifically bound, as opposed to round eight. Among the selected nine sequences characterized by
bioinformatics analysis and ELONA, an aptamer sequence with the highest specificity and affinity
for the target protein was identified and further examined. Aptamer sequence (6) was screened in a
concentration-dependent assay, specificity analysis was performed, and its potential secondary and
tertiary structures were predicted, which enabled us to test one of the possible putative interactions
with CEA. Finally, aptamer sequence (6) labelled with a Cy5 fluorescent tag was used in confocal
microscopy to observe its binding towards the CEA expressed in HT-29 human colon adenocarcinoma
cell line
Description
Keywords
Type of access: Open Access, aptamer, carcinoembryonic antigen (CEA), selection, SELEX, ELONA, bioinformatics analysis, HT-29 human colon adenocarcinoma
Citation
Yunussova, N., Sypabekova, M., Zhumabekova, Z., Matkarimov, B., & Kanayeva, D. (2022). A Novel ssDNA Aptamer Targeting Carcinoembryonic Antigen: Selection and Characterization. Biology, 11(10), 1540. https://doi.org/10.3390/biology11101540