1. Design Principles

Selection of Target Gene Sequence: The target gene sequence is known, and the antisense oligonucleotide, typically 15-30 bases in length, has a simple structure that makes it easy to design and synthesize in large quantitiesin vitro. Antisense nucleic acids are independent of the cell cycle and can enter both proliferating and non-proliferating cells. Antisense oligonucleotides do not contain viral sequences, so they do not trigger immune responses or integrate into host chromosomes. Antisense oligonucleotides targeting mRNA are ideal because mRNA accounts for only 2%-5% of total RNA in cells, making its copy number relatively low. These oligonucleotides can directly inhibit the translation process, with the most effective targeting occurring at the translation initiation region.

Length and Base Composition: The optimal length of an antisense oligonucleotide is typically considered to be between 11 and 25 bases, with 15-20 bases being the most common length. Sequences that are too long are not easily absorbed by cells, while sequences that are too short lack specificity. The G and C content should account for 60%-65% of the total bases in the sequence. High G and C content can cause the antisense oligonucleotide to "aggregate," reducing specificity, while low G and C content may decrease the affinity between the antisense oligonucleotide and the target sequence.

Avoiding Self-Pairing: When designing the sequence, it's essential to avoid self-pairing of the bases in the antisense oligonucleotide. Self-pairing can lead to the formation of secondary structures, which would hinder the oligonucleotide's ability to effectively bind to the target sequence.

2. Synthesis and Purification

Synthesis Methods: Antisense oligonucleotides can be synthesized using chemical synthesis methods and other techniques.  

Purification: Once synthesized, antisense oligonucleotides must undergo purification to ensure their quality and concentration. The purification process eliminates impurities, improving the overall purity of the oligonucleotides.

3. Verification and Modification

Experimental Validation: Each designed antisense oligonucleotide should be experimentally validated to confirm its specificity and evaluate its inhibitory effectiveness.

Modification and Optimization: Based on experimental results, the design of the antisense oligonucleotides must be modified and optimized to improve their specificity and inhibitory effectiveness.

What Are the Applications of Synthesized Antisense Oligonucleotides?

Regulation of Gene Expression: Antisense oligonucleotides utilize base pairing principles to specifically bind to the DNA or mRNA of target genes, disrupting processes such as gene unwinding, replication, transcription, mRNA splicing, processing, and translation. This binding inhibits target gene expression, thereby regulating key biological processes like cell growth and differentiation.

Disease Treatment: Antisense oligonucleotides can be used to treat certain hereditary diseases by inhibiting the expression of mutated genes to alleviate symptoms. Additionally, ASOs can be used in cancer therapy to inhibit the growth and spread of tumors by suppressing the expression of tumor-related genes.

Drug Development: As a new class of molecular drug, antisense oligonucleotides are highly specific and targeted in their action. This makes them of significant application value in drug development. By designing and synthesizing ASOs for specific target genes, new drugs with high efficiency, low toxicity, and low side effects can be developed.

Biomedical Research: By inhibiting the expression of a specific gene to study its role in biological processes, antisense oligonucleotides are an invaluable tool in gene function research.  ASOs offer promising new strategies and approaches for advancing gene therapy research.

Synbio Technologies |Antisense Oligonucleotides

At Synbio Technologies, we proudly provide high quality antisense oligonucleotides with maximum binding affinity and stability through optimized synthesis processes. Various ASO modification types are available, while the cutting-edge methods and advanced technologies, such as high-performance liquid chromatography (HPLC) purification and electrospray ionization mass spectrometry (ESI-MS), ensure the highest level of purity and accuracy. We are experts in empowering nucleic acid drug research and development.