Oligonucleotides are short chains of nucleotides, typically up to 20 linked units, that can be formed either enzymatically through the cleavage of larger molecules or via targeted chemical synthesis. In chemical synthesis, oligonucleotide synthesis is the process by which nucleoside phosphoramidites—key building blocks of oligonucleotides—are synthesized. These phosphoramidites are derivatives of natural or synthetic nucleosides. Nucleotides, which consist of a nucleoside covalently bonded to a phosphate group, are linked in a specific sequence to create the desired oligonucleotide product. Due to their relatively small size, oligonucleotides are considered small biopharmaceutical molecules.

1.How Are Oligonucleotides Used?

Research, Diagnostic, Forensic, and Therapeutic Applications

Synthetic oligonucleotides are most commonly used as short primers and probes (up to 30 nucleotides long) in various applications. As primers, oligonucleotides initiate enzymatic reactions to create numerous copies of a target DNA or RNA sequence, as seen in techniques like polymerase chain reaction (PCR) and Sanger sequencing. These primers are essential in DNA sequencing, gene expression studies, cloning, and molecular diagnostics.

As probes, oligonucleotides bind to specific DNA or RNA sequences to detect their presence in a sample. This function is utilized in blotting techniques (such as northern and southern blotting), microarrays for gene expression analysis, and molecular diagnostics for detecting genetic diseases or identifying pathogens.

2.Oligo Therapeutics/Gene Therapy

In therapeutic applications, antisense oligonucleotides (ASOs), typically 20 to 30 nucleotides long, are used to inhibit or silence undesirable RNA sequences. This process can block the expression of harmful proteins associated with certain diseases. Research in oligonucleotide-based therapeutics has grown significantly, leading to the approval of several ASO drugs in recent years.

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