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Aptamer Selection and Aptamer Sequencing
Aptamer Selection and Aptamer Sequencing
The process of SELEX depicted in a figure made with Biorender.
Systematic Evolution of Ligands by Exponential Enrichment (SELEX)
Systematic Evolution of Ligands by Exponential Enrichment (SELEX)
SELEX is the process performed in order to find the sequences of aptamers with the most binding affinity to the target protein. A pool of single-stranded DNA strands or RNA strands are incubated with the target (ex. protein or small molecule), then unbound nucleic acids are washed away from the target while the bound nucleic acids are eluted and amplified with PCR. After multiple rounds of selection, the selection pools can be sent for sequencing and the sequencing data can be analyzed to identify top aptamer candidates.
Keywords
Keywords
Binding Affinity
Binding Affinity
The strength of the binding interaction between a biomolecule and its binding partner. The stronger the binding affinity between the aptamer and target, the better.
Aptamer
Aptamer
Aptamers are oligonucleotide sequences that bind to a specific target molecule. They can be selected for and sequenced. Aptamers are usually created by selecting them from a large starting library. They have a wide variety of possible uses. They are used in things like diagnostics and therapeutics.
Target Ligand
Target Ligand
The target molecule (protein, carbohydrates, toxins, etc) which one is trying to find the corresponding aptamer to.
Relevant Papers
Relevant Papers
Aptamers as therapeutic and diagnostic agents.
Aptamers as therapeutic and diagnostic agents.
Brody, E. N., & Gold, L. (2000). Aptamers as therapeutic and diagnostic agents. Journal of biotechnology, 74(1), 5–13. https://doi.org/10.1016/s1389-0352(99)00004-5
This article describes how aptamers can be modified to work more efficiently. The 2'OH of the ribose groups are changed to 2'F or 2'NH2 groups so the aptamers work better at reaching their target in animals and humans. By increasing the molecular weights of the aptamers, they are able to the prevent degradation of the aptamers in blood by nucleases.
Development of DNA aptamers using Cell-SELEX.
Development of DNA aptamers using Cell-SELEX.
Sefah, K., Shangguan, D., Xiong, X., O'Donoghue, M. B., & Tan, W. (2010). Development of DNA aptamers using Cell-SELEX. Nature Protocols, 5(6), 1169-1185. doi:doi:10.1038/nprot.2010.66
The authors of the article describe the process of SELEX and how the aptamers produced can bind to the cell of interest. The single stranded DNA strands are incubated with the target cells, and nonbinding strands are washed away while the binding strands are incubated. This leads to an enriched pool of strands of DNA with affinity to the target cell, which are then amplified with PCR. This whole process takes around 3 months.
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