Development and characterization of DNA aptamer against Retinoblastoma by Cell-SELEX

Retinoblastoma (RB) is the most common paediatric intraocular tumour. The management of RB has improved the survival and vision with recent advances in the treatment. Improved therapeutic approaches focussing on targeting tumours and minimizing the treatment-associated side effects are being develop...

Full description

Saved in:
Bibliographic Details
Published in:Scientific reports 2022-09, Vol.12 (1), p.16178-16178, Article 16178
Main Authors: Maradani, Bhavani Shankar, Parameswaran, Sowmya, Subramanian, Krishnakumar
Format: Article
Language:English
Subjects:
631/67/2332
692/699/67/1484
692/700/139
Affinity
Aptamers
Cancer
Cerebrospinal fluid
Flow cytometry
Glial cells
Humanities and Social Sciences
Mueller cells
multidisciplinary
Next-generation sequencing
Phylogeny
Proteinase
Retina
Retinoblastoma
Science
Science (multidisciplinary)
Tumors
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Retinoblastoma (RB) is the most common paediatric intraocular tumour. The management of RB has improved the survival and vision with recent advances in the treatment. Improved therapeutic approaches focussing on targeting tumours and minimizing the treatment-associated side effects are being developed. In this study, we generated a ssDNA aptamer against RB by cell-SELEX and high-throughput sequencing using Weri-RB1 cell line as the target, and Muller glial cell line Mio-M1 as the control. Three aptamers were selected based on the number of repetitions in NGS and phylogenetic relationship and evaluated by flow cytometry to assess their binding affinity and selectivity. The dissociation constant, Kd values of three selected aptamers were found to be in the nanomolar range. Aptamer VRF-CSRB-01 with the best binding affinity and a Kd value of 49.41 ± 7.87 nM was further characterized. The proteinase and temperature treatment indicated that VRF-CSRB-01 targets surface proteins, and has a good binding affinity and excellent selectivity under physiological conditions. The aptamer VRF-CSRB-01 was stable over 72 h in serum and 96 h in cerebral spinal fluid and vitreous. With the high affinity, specificity, stability and specific recognition of clinical RB tumours, VRF-CSRB-01 aptamer holds potential for application in diagnosis and targeting RB.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-022-20660-3