The CRISPR-Cas9 system has rapidly been adopted for many genome engineering
applications including creating gene knock-outs/knock-ins, genome mutagenesis,
and gene activation and inhibition studies. This Application Note demonstrates
how several Agilent products can be used in a CRISPR-Cas9 workflow, starting
with the design of a guide RNA (gRNA) against a specific target, synthesis of
the gRNA, quality control methods to ensure the integrity of the gRNA, and a
Cas9 nuclease activity assay to test the efficacy of the generated gRNA. gRNAs
were designed and synthesized against two target genes: ZFP42 and GUSB. The
synthesized gRNAs were assessed for their overall quality and cleavage efficacy
using the Agilent 2100 Bioanalyzer system
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