Systematic evolution of ligands by exponential enrichment
Template:Infobox laboratory technique
Systematic evolution of ligands by exponential enrichment (SELEX) is an in vitro selection technique used to identify nucleic acid ligands, known as aptamers, that have a high affinity for specific target molecules. This method is widely used in molecular biology, biochemistry, and biotechnology for the development of diagnostic and therapeutic agents.
History[edit | edit source]
The SELEX process was independently developed by two research groups in 1990. The technique has since become a fundamental tool in the field of molecular evolution and has been adapted for various applications, including the development of biosensors, drug delivery systems, and targeted therapies.
Process[edit | edit source]
The SELEX process involves several iterative steps:
- **Library Preparation**: A large library of random oligonucleotides is synthesized. These oligonucleotides can be composed of DNA, RNA, or modified nucleic acids.
- **Binding**: The library is incubated with the target molecule, allowing potential aptamers to bind to the target.
- **Partitioning**: Bound oligonucleotides are separated from unbound ones. This can be achieved through various methods such as affinity chromatography, filtration, or magnetic separation.
- **Amplification**: The bound oligonucleotides are amplified using polymerase chain reaction (PCR) for DNA or reverse transcription followed by PCR for RNA.
- **Iteration**: The amplified oligonucleotides are subjected to additional rounds of binding, partitioning, and amplification to enrich the pool for high-affinity aptamers.
Applications[edit | edit source]
SELEX has numerous applications in both research and clinical settings:
- **Diagnostics**: Aptamers selected through SELEX can be used in biosensors for the detection of various biomarkers.
- **Therapeutics**: Aptamers can be developed as therapeutic agents to target specific proteins or cells in diseases such as cancer and viral infections.
- **Research Tools**: Aptamers can be used to study protein-protein interactions, cell signaling pathways, and other biological processes.
Advantages[edit | edit source]
SELEX offers several advantages over traditional methods of ligand discovery:
- **Specificity**: Aptamers can be selected to bind with high specificity to their target molecules.
- **Versatility**: The SELEX process can be adapted to select aptamers for a wide range of targets, including small molecules, proteins, and even whole cells.
- **Stability**: Aptamers are generally more stable than antibodies and can be easily modified to enhance their properties.
Limitations[edit | edit source]
Despite its advantages, SELEX also has some limitations:
- **Complexity**: The SELEX process can be time-consuming and requires specialized equipment and expertise.
- **Cost**: The synthesis and amplification of large oligonucleotide libraries can be expensive.
- **Target Accessibility**: Some targets may be difficult to access or present in low abundance, complicating the selection process.
See also[edit | edit source]
References[edit | edit source]
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Contributors: Prab R. Tumpati, MD