SpCas9 Nuclease

For genome editing

  • High genome editing efficiency with jetCRISPR™
  • Especially designed for transfection
  • Minimize off target-effects using RNP approach
  • Better Cas9 activity control by using RNP

Specifications

Product

SpCas9 Nuclease

Application

Genome editing (RNP delivery)

Sequence

NLS-SpCas9-NLS

Concentration

Solution at 10 µg/µL

Molecular Weight

162 kDa

Storage

-20 °C

Summary

Successful delivery of the gRNA and the Cas9 into cells is essential to guarantee the high genome editing efficiency. The most efficient CRISPR-Cas9 method is based on the direct delivery of pre-complexed gRNA and Cas9 protein, referred to as Ribonucleoprotein (RNP) delivery.

The sequence of our SpCas9 nuclease is carefully optimized to improve its nuclear import, nuclease editing efficiency, and to reduce off-targeting effects. Our SpCas9 nuclease has been developed to reach the highest genome editing efficiency with our RNP transfection reagent jetCRIPSR™.

Combine the leading technology for RNP delivery using jetCRISPR™ with the optimal Cas9 protein structure to introduce specific Cas9-mediated genome editing efficiency on a wide variety of cell types.

Read more about jetCRISPR™

Ordering information

ProductReference NumberProduct size
SpCas9 Nuclease722-100100 µg
jetCRISPR™502-010.1 ml
jetCRISPR™502-070.75 ml
jetCRISPR™502-151.5 ml

Optimized sequence

Polyplus-transfection’s SpCas9 Nuclease is a recombinant S. pyogenes Cas9 nuclease that has been optimized to improve nuclear entry following jetCRISPR™-mediated intracellular delivery. The sequence of spCas9 nulcease contains a single SV40 nuclear localization sequence (NLS) in N- and C-terminus (Fig. 1).

SpCas9 Nuclease - Design
Fig. 1: Optimized SpCas9 Nuclease design for efficient genome editing.

High genome editing efficiency

jetCRISPR™ is an innovative transfection reagent for the delivery of RNP complexes to achieve CRISPR-Cas9 mediated genome editing. Choosing the optimal Cas9 sequence is key in enhancing targeted nuclear delivery of the Cas9 protein with the gRNA. Polyplus-transfection’s SpCas9 nuclease leads to higher genome editing than other commercially available Cas9 proteins using jetCRISPR™ transfection reagent (Fig. 2).

Spcas9 Nuclease - Cas9 comparison using jetCRISPR
Fig. 2: Higher genome editing obtained with SpCas9 Nuclease compared to other Cas9 Proteins. RNP transfections were performed in HEK-293 cells using 30 nM RNP (HPRT1 sgRNA + Cas9 protein) with 0.3 µl of jetCRISPR™ reagent per well of 96-well plate. At 48 h post-transfection, genome editing was assessed by calculating the percentage (%) of INDEL using the T7 endonuclease method. The INDEL % was determined by using Genetools software (Syngene®).

jetCRISPR™ transfection reagent achieves higher genome editing efficiency than other competitors, which makes it the product of choice for CRISPR applications.

jetCRISPR - Comparison of genome editing efficiency
Fig. 2: Superior genome editing efficiency obtained with jetCRISPR™ in comparison with Lipofectamine® CRISPRMAX™. RNP transfections were performed in A549 and HEK-293 cells using 30 nM RNP (SpCas9 Nuclease and HPRT1 sgRNA) with 0.3 µl of jetCRISPR™ reagent or 0.3 µl of Lipofectamine® CRISPRMAX™, per well of a 96-well plate. At 48 h post-transfection, genome editing was assessed by calculating the percentage (%) of INDEL using the T7 endonuclease method. The INDEL % was determined by using Genetools software (Syngene®).

Video

Alengo Nyamay’antu (PhD), Technical Support Specialist, talks about transfection trends in genome editing. She provides an overview of solutions to delivery both guide RNA and Cas9 nuclease into cells.

FAQ

If you have any questions regarding SpCas9 Nuclease and jetCRISPR™, please visit our dedicated Frequently asked questions or contact us at support@polyplus-transfection.com.

We provide a full range of reagents for all your CRISPR experiments (Table 1 & Fig. 4):

Polyplus-transfection - Table CRISPR reagents
Table 1: Range of transfection reagents for CRISPR experiments

Fig. 6: Polyplus-transfection - Product range for CRSIPR experiment
Fig 4: Range of in vitro transfection reagents for CRISPR experiments

Every batch of SpCas9 nuclease is tested by an in vitro activity assay using gRNA and linearized DNA target. Each vial is provided with a Certificate of Analysis.