Application of NGS-Based Technology in sgRNA Sequencing QC

Read our new application note to discover how GenScript leverages new NGS-based technology to ensures cGMP sgRNA sequence quality.

Gene editing therapies powered by CRISPR/Cas9 tools continue to advance, enabling innovative and often curative solutions to various diseases. Investigators developing cell and gene therapies with critical components, such as Cas9 nuclease and single guide RNA (sgRNA), must navigate a challenging landscape to meet the dynamic regulatory requirements pertinent to these novel biological products. Therefore, it is essential to leverage QC methods that can demonstrate sufficient sensitivity and specificity for gene editing components used in studies to expedite IND submission.

Widely adopted solid-phase oligo synthesis strategy for sgRNA involves several steps including a capping step which could lead to the generation of G->A single nucleotide mutations, leading to increased risk for off-target editing. Yet commonly used analytical methods, including HPLC and high-resolution MS fail to differentiate this type of mutation. To ensure sgRNA sequence fidelity, especially at cGMP grade, GenScript developed our own proprietary NGS based approach in sequencing through the sgRNA with high sensitivity. Read our application note to learn more.