Antisense Oligonucleotide (ASO) Off-Target Detection and Analysis Service
Introduction
ASO drugs target pre-mRNA or mature mRNA and induce degradation to achieve therapeutic effects. However, due to sequence similarities between target mRNA and non-targeted mRNA, mismatch, insertion, and deletion can occur, leading to off-target effects, triggering unexpected genetic mutations or functional changes that affect study results or therapeutic efficacy, and may raise safety concerns.
Therefore, in the development process, it is important to understand the off-target effects of ASO drugs as early as possible. Through professional off-target effect detection and analysis services, drug developers can identify potential safety risks early in drug discovery and provide safer and more effective treatment options for patients.
Fig.1 Possible mechanism of off-target effect.
Commonly Used Off-target Effect Detection Method
Whole genome sequencing (WGS)
WGS is one of the simplest and most effective ways to detect off-target effects. By sequencing the entire genome, a wide range of mutations resulting from gene editing were comprehensively detected, including single nucleotide mutations (SNV), insertion-deletion variants (Indels), chromosome rearrangements, copy number variants (CNV), and structural variants (SV). This method is highly sensitive and comprehensive and can detect very off-target events of low frequency, but the cost is relatively high, the data analysis process is complex and requires the support of a professional bioinformatics team.
Fig.2 Whole genome sequencing diagram.Distributed under CC BY-SA 2.5, from Wiki, without modification.
Target capture and sequencing
Target acquisition sequencing is a method for high-throughput sequencing of specific regions, and the acquisition probe is designed according to the predicted off-target sites for targeted sequencing. Compared with whole genome sequencing, this method has a lower cost and can detect specific off-target sites, improving the pertinence and accuracy of detection. However, coverage is limited and it may not be possible to detect all potential off-target sites.
Digenome-seq
Digenome-seq uses in vitro cutting technology to simulate the gene editing process, and sequencing the genomic DNA after cutting, comparing the genomic sequences before and after cutting, can identify off-target effects. The operation is simple and the cost is relatively low, but it may be affected by the efficiency and specificity of in vitro cutting.
PCR-based detection method
Specific endonucleases are used to cut PCR products, and then off-target sites are identified by analyzing the sequence of the cut products. The operation is simple and low-cost, but it may be affected by the amplification efficiency and specificity of PCR, and only off-target effects can be detected at specific locations.
The Highlight of Our Service
Creative Biolabs offers a comprehensive range of ASO drug off-target testing services, including:
Target screening and validation: Use advanced bioinformatics tools to screen and identify potential sequences on the human genome that overlap with ASO or target genes to ensure accuracy.
Off-target effect evaluation: Comprehensive evaluation of off-target effects of drug candidates through functional screening methods. Creative Biolabs focuses on the silencing effect of drugs on non-target genes, as well as possible biological effects and adverse reactions.
Reporting and consulting: Detailed presentation of test results and data analysis. The team of experts will provide professional advice and recommendations to help customers develop subsequent research and development strategies.
Comprehensive coverage: Creative Biolabs' services cover the whole process from target screening to off-target effect assessment, ensuring full and accurate identification of potential risks.
Professional team: Creative Biolabs has a team of experienced experts with a deep background in biology, able to provide customers with professional technical support and solutions.
Efficient and fast: Creative Biolabs uses advanced testing technology and processes to ensure high efficiency and accuracy of test results. At the same time, we provide flexible sample delivery methods and diversified payment channels for the convenience of customers.
Reliability of data: All targets are tested using functional methods, which can reduce false negatives and avoid safety risks in the later stage to a greater extent.
Through its professional off-target effect detection services, Creative Biolabs helps drug developers detect off-target effects of ASO drugs as early as possible and reduce potential risks, ensuring their safety and efficacy. If you are developing an ASO drug and would like to learn more about the detection of off-target effects, please contact us.
