High efficiency, RNA-guided
- All-in-one Cas9/gRNA plasmids
- Injection-ready Cas9 mRNA
- Packaged Cas9 lentivirus
- Multiplex gRNA cloning kits
- Expert technical support
Cas9 Synthetic mRNA and T7 gRNA Production
The type II prokaryotic CRISPR (clustered regularly interspaced short palindromic repeats) adaptive immune system has been shown to facilitate RNA-guided sequence-specific DNA cleavage, which provides a new class of genome engineering tools. To make the RNA-directed Cas9 system more efficient, and convenient to use for in vivo applications, SBI has developed a CRISPR/Cas9 mRNA system, which includes functionally-validated Cas9 mRNA (cat# CAS500A-1), T7 gRNA cloning vector and T7 gRNA producing kit. To avoid reconstituting the CRISPR/Cas9 RNA processing machinery, a custom gRNA (crRNA-tracrRNA chimeric transcript) can be generated from the ready-to-use linearized T7 gRNA cloning vector (cat# CAS510A-1) through the use of annealed oligonucleotide duplexes encoding the 20bp target sequence upstream of PAM (NGG). As the custom gRNA is under the control of T7 promoter, it is ready for in vitro transcription (IVT) with the T7 gRNA synthesis kit (cat# CAS510A-KIT). The AAVS1 gRNA sequence was cloned into the CAS510A-1 T7 vector and scaffolded-gRNA targeting the AAVS1 site was generated in vitro. These gRNAs were co-transfected with the Cas9 synthetic mRNA in combination with the AAVS1 HR vector harboring a GFP marker. The activities of the Cas9 mRNA + AAVS1 gRNA transfection was compared with that of the EF1 Cas9 SmartNuclease-AAVS1 gRNA all-in-one vector system. Cells were imaged for GFP fluorescence after 3 days.
Rapidly Generate Mouse Models using Cas9 SmartNickase
Read the new case study about a recent publication from UC Davis' Mouse Biology Program in which injection-ready Cas9 nickase mRNA and gRNA were used to flox a target gene and rapidly generate a mouse model with a conditional KO allele. SBI's Cas9 SmartNickase mRNA and gRNA IVT Synthesis kits are highlighted.
Dr. Angus Lee