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CRISPR/Cas9 SmartNucleases™

High efficiency, RNA-guided
genome editing

CRISPR/Cas9 Technologies

  • All-in-one Cas9/gRNA plasmids
  • Injection-ready Cas9 mRNA
  • Packaged Cas9 lentivirus
  • Multiplex gRNA cloning kits
  • Expert technical support

PrecisionX™ Multiplex gRNA Cloning Kits

Create CRISPR/Cas9 constructs with multiple gRNAs simultaneously for more precise genome editing

SBI has developed a revolutionary cloning kit in order to facilitate the cloning of gRNAs into CRISPR/Cas9 vectors, the PrecisionX Multiplex gRNA Cloning Kit (Cat# CAS9-GRNA-KIT). This system allows for the cloning of multiple gRNAs into any Cas9/gRNA all-in-one expression vector or gRNA cloning vector of the customer's choice, including SBI's PrecisionX Cas9/gRNA SmartNuclease plasmids and lentivectors. The Multiplex gRNA Cloning Kit is also compatible with the most popular Cas9/gRNA cloning vectors developed in labs across the world, such as pX330, pX335, pX458, and pX459.

How the Multiplex gRNA Cloning Kit Works

   

The PrecisionX multiplex gRNA cloning kit provides the H1 and U6 promoter blocks to easily build multiple gRNA cassettes. In step 1, primers with overlapping ends containing the the desired guide RNA (designed by user) and the scaffold-promoter block (from the kit) are combined in a PCR reaction to generate the PCR amplicon containing both gRNAs. Secondly, the PCR amplicon created in step 1 is combined with the fusion reaction mix along with the linearized destination expression vector for seamless construction. The cloning process is extremely efficient and the kit can be adapted to two or more gRNAs in a single fusion reaction. A schematic of the assembly workflow is shown to the right.