Recent advances in histopathology reporting 2, tumor sub-type identification 3, 4 and cell-free DNA diagnostics 5, 6 have improved the prognosis of RB. However, advances in imaging, local forms of therapy, and systemic chemotherapy have improved outcomes in better survival and vision salvage. Retinoblastoma is highly aggressive and leads to intra-orbital, intracranial, and even systemic metastasis. Retinoblastoma (RB) is the most common primary intraocular malignant tumour in children, with an incidence of 1:16,000 to 18,000 live births worldwide 1. 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. The aptamer VRF-CSRB-01 was stable over 72 h in serum and 96 h in cerebral spinal fluid and vitreous. 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. Aptamer VRF-CSRB-01 with the best binding affinity and a Kd value of 49.41 ± 7.87 nM was further characterized. The dissociation constant, Kd values of three selected aptamers were found to be in the nanomolar range. 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. 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. Improved therapeutic approaches focussing on targeting tumours and minimizing the treatment-associated side effects are being developed. The management of RB has improved the survival and vision with recent advances in the treatment. Retinoblastoma (RB) is the most common paediatric intraocular tumour.
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