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ASO In Vitro Screening Service


An antisense oligonucleotide (ASO) is a short strand of deoxyribonucleotide analogue that hybridizes with the complementary mRNA in a sequence-specific manner via Watson-Crick base pairing. To be useful as a drug, ASO must be much more stable than ordinary. Creative Biolabs has extensive experience with various antisense technologies to modulate RNA expression. Our sophisticated technologies, professional technicians are available to provide ASO in vitro screening service for worldwide customers.

Action Mechanisms of Antisense Oligonucleotides (ASOs)

ASO is not only a useful experimental tool in protein target identification and validation, but also a highly selective therapeutic strategy for diseases with dysregulated protein expression. Antisense oligonucleotide-induced protein knockdown is usually achieved by the induction of RNase H endonuclease activity that cleaves the RNA-DNA heteroduplex. This results in the degradation of target mRNA while leaving the ASO intact. Other ASO mechanisms include translational arrest by steric hindrance of ribosomal activity, interference with mRNA maturation by inhibiting splicing and destabilization of pre-mRNA in the nucleus. These modes of action of antisense oligonucleotides are shown in Figure 1.

Modes of action of antisense oligonucleotides. Figure 1. Modes of action of antisense oligonucleotides. (Chan, 2006)

ASOs Efficacy and Specificity

In terms of experimental and therapeutic applications of ASO, there are two important factors should be considered: efficiency and specificity. Previous studies have indicated that ASOs can discriminate differences in target RNAs as small as a single base change. Recently, some studies have shown that oligonucleotides can have biological effects that are not attributable to specific degradation or blockade of their target RNAs. These influences can result from sequence-specific aptameric effects of oligonucleotides, non-sequence specific binding of oligonucleotides to proteins, or RNA cleavage due to partial sequence matches. Therefore, it is important to analyze the efficiency and specificity of ASOs.

ASOs Efficacy Screening

In the absence of efficient design algorithms, efficient ASOs against the RNA target of interest are designed via experimental screening. Based on the fully optimal high-throughput workflow, Creative Biolabs can simultaneously test thousands of ASOs in vitro and finally select expected candidate ASOs. This process finally selects candidate ASOs that induce knockdown of the target RNA in a dose-responsive manner.

ASO Specificity Analysis

ASOs also carries the risk of causing unintended toxicities and/or side effects. This is necessary and crucial to know potential off-target related toxicity and side effects. Creative Biolabs allows you to further screens the candidate ASOs by the advanced assessment (in silico assessment) and experimental validation of putative off-target related toxicity and side effects. This experimental validation platform can map the off-target effects of candidate ASOs and list validated off-target genes for each candidate ASO.

Creative Biolabs’ advanced technologies and highly experienced staff are committed to advancing your program with our ASO in vitro screening services and reducing the overall development timeline. If you have any questions in the ASO in vitro screening services, please don’t hesitate to contact us for more information.

References

  1. Chan, J.; et al. (2006). Antisense oligonucleotides: from design to therapeutic application. Clinical and experimental pharmacology and physiology. 33(5-6): 533-540.

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