AID-Seq    

Creative Biogene CRISPR/Cas9 Platform provides AID-Seq service for customers to assess off-target effect of CRISPR/Cas9. With years of experience in CRISPR-Cas9 and NGS field, our talented scientists will work closely with you to assist you in off-target effects analysis via AID-seq.

Engineered nucleases, which can introduce targeted DNA double-stranded breaks into the genome of living cells, are widely used as genome-editing tools. However, their clinical applications are limited due to the off-target issue. The undesired off-target mutations potentially lead to unintended effects in research and clinical applications. Thus, sensitive and unbiased genome-wide methods are developed to understand the genome locations and frequency of nuclease-induced DSB activity.

AID-seq (adaptor-mediated off-target identification by sequencing) is a novel and highly sensitive method to identify genome-wide off-target mutations. This method enables to detect the low-frequency off-target sites, while minimizing the background noise and reducing the false positive rate. Moreover, this method provides a possibility to assess hundreds of sgRNA in the same time by the pooled strategy.

Figure 1. Schematic overview of AID-Seq method¹

AID-seq is an NGS-based method to detect off-target cleavage of CRISPR/Cas9. This method is charactered by using two different adaptors, hairpin i7 adaptor and biotin i5 adaptor. Genome DNA extracted from samples is randomly sheared and ligated with hairpin i7 adaptor. The double-tailed DNA fragments are further digested by Cas9 or Cas12a RNP complexes and then ligated with biotin i5 adaptor. These biotin adaptor ligated DNA will be enriched by streptavidin beads and analyzed by next-generation sequencing.

As a professional provider, Creative Biogene CRISPR/Cas9 Platform employs various approaches to detect the off-target cleavages of CRISPR/Cas9, like AID-Seq. This service covers from sample preparation, sequencing to data analysis. Once project finishes, reliable report and raw data will be delivered to customers.

For more information, please contact us. We are waiting for your interest projects.

Related Services

• CRISPR Off-Target Effects Analysis
• PEM-Seq
• Target Region Sequencing
• SgRNA Design and Confirmation