Tag-seq

Tag-seq enables genome-wide identification and profiling of DNA double-strand breaks (DSBs) induced by CRISPR-associated protein (Cas) nucleases.

Key Features:

• Efficient off-target detection: Detects off-target events caused by Cas nucleases, including SpCas9 variants and Cas12a/Cpf1, via integration of an optimized double-stranded oligodeoxynucleotide sequence called "Tag".
• Next-generation sequencing integration: Leverages next-generation sequencing (NGS) to trace integrated Tag sequences and profile nuclease-induced DSBs genome-wide.
• Optimized library preparation: Employs a one-step library preparation procedure to capture Tag-containing fragments for sequencing.
• Applicability to transposon mapping: Identifies integration sites of exogenous genes introduced by the Sleeping Beauty transposon system.

Scientific Applications:

• CRISPR/Cas specificity assessment: Genome-wide evaluation of on- and off-target cleavage by Cas nucleases to inform precision and safety of genome editing.
• Integration site identification: Mapping of exogenous gene insertion sites from systems such as the Sleeping Beauty transposon.

Methodology:

Traces integrated Tag sequences from next-generation sequencing (NGS) reads and uses a one-step library preparation to capture Tag-containing fragments.