jetCRISPR™

RNP transfection reagent for genome editing

  • Specifically designed for Cas9 protein and guide RNA delivery
  • High genome editing efficiency
  • Excellent cell viability and morphology
  • Fast and reliable gene editing
  • Easy to use: Reverse and Forward protocols

Specifications

Reagent

jetCRISPR™

Molecule delivered

Ribonucleoprotein: Guide RNA and Cas9 protein

Application

CRISPR mediated Genome editing

Cell types

Adherent and suspension cells

Number of transfections

1.5 ml of jetCRISPR™ transfection reagent is sufficient to perform up to 1 250 transfections in 24-well plates or 300 transfections in 6-well plates

Storage

4 °C, stable for 6 months (502-01) to at least one year (other packaging sizes) when stored appropriately

Summary

The CRISPR/Cas9 engineered nuclease system is a powerful and highly specific genome editing tool. CRISPR/Cas9 is a two component system with a guide RNA (gRNA) molecule that drives the Cas9 nuclease to a specific targeted sequence within the genome in order to introduce genetic modifications (mutations, insertions or deletions).

jetCRISPR™ is an innovative reagent especially designed to deliver RNP in a CRISPR/Cas9 experiment. jetCRISPR™ leads to high CRISPR genome editing efficiency and keep an excellent cell viability and morphology. Use the leading technology for CRISPR/Cas9 RNP delivery!

Protocols and Files

Ordering information

Catalog NumberAmount of reagent
502-010.1 ml
502-070.75 ml
502-151.5 ml

1.5 ml of jetCRISPR™ transfection reagent is sufficient to perform ca. 300 transfections in 6-well plates. Bulk quantities are available upon request, please contact us for more details.

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High Genome Editing efficiency

jetCRISPR™ has been specifically designed for high transfection efficiency of RNP (guide RNA and Cas9 protein complex), thus leading to a high cas9-mediated genome editing efficiency on a wide variety of targets in different cell types (Fig. 1 & 2).

jetCRISPR - Genome editing efficiency
Fig. 1: High genome editing efficiency using jetCRISPR™ in HeLa cells. RNP transfections were performed in HeLa cells using several RNP concentrations of Cas9 protein and HPRT1 sgRNA or negative control in combination with 0.3 µl of jetCRISPR™ reagent per well of a 96-well plate. At 48 h post-transfection, T7 digestion products were run on a 2% agarose gel and stained with BET displayed by G:Box transilluminator (Syngene®). Acquisitions were carried out with the Genesnap software (Syngene®) and INDEL quantifications were performed with the Genetools software (Syngene®). 1: Uncleaved fragment of 1083 bp, 2: long cleaved fragment of 827 bp, 3: short cleaved fragment of 256 bp.

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 (Cas9 Protein and HPRT1 sgRNA) with 0.3 µl of jetCRISPR™ reagent or 0.3 µl of Lipofectamine® CRISPRMAX™, 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®).

Excellent cell viability

At Polyplus-transfection, we are proud to work with reagents that are extremely gentle to cells, maintaining an excellent cell viability and morphology throughout the whole duration of the experiments. jetCRISPR™ is compatible with both serum and antibiotics, hence RNP transfection can be performed in the optimal conditions for healthy cells (Fig. 3).

jetCRISPR - Cell Viability
Fig. 3: Excellent cell viability and morphology at 48 h after transfection with jetCRISPR™. RNP transfections were performed on HEK-293 and A549 cells using 30 nM RNP (Cas9 Protein and HPRT1 sgRNA) with 0.3 µl of jetCRISPR™ reagent per well of 96-well plate. At 48 h post-transfection, the cell morphology was visualized by phase contrast ZOE Fluorescent Cell Imager (BIORAD®).

Fast and Reliable gene editing

Delivering the Cas9 directly as a protein instead as a plasmid allows for faster genome editing as the protein is readily available for use. Using jetCRISPR™ to deliver Cas9 protein and gRNA leads to fast and reliable gene editing (Fig. 4). In addition, the Cas9 protein will be cleared from the cells faster than the plasmid, hence minimizing off-target effects and leading to more specific genome editing (Kim S, et al; Genome Research 2014; 24:1012–1019).

jetCRISPR - Fast and reliable gene editing
Fig. 4: Fast and reliable gene editing efficiency obtained with jetCRISPR™ in HEK-293 cells. RNP transfections were performed in HEK-293 cells using 30 nM RNP (Cas9 Protein and HPRT1 sgRNA) 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®).

Easy to use

jetCRISPR™ is provided as a ready to use solution. Transfecting RNP is now a very straightforward process only requiring a few steps: form the RNP complexes, add the transfection reagent and add to the wells!

Both reverse and forward transfections are perfectly fine to use with jetCRISPR™, hence allowing you to adapt your protocol to suit the cells that you are working with, the plate format as well as the time of analysis. By using the reverse protocol (Fig. 5), data will be available a day earlier than if you use the forward protocol.

jejetCRISPR protocol - Reverse
Fig. 5: jetCRISPR™ reverse protocol

 

Successful delivery of the gRNA and the Cas9 into cells is indispensable to guarantee the high genome editing efficiency that is required to generate new cell or animal models. While DNA-free delivery systems in which the Cas9 protein is delivered as a protein/gRNA (RNP) have become attractive alternative, as they overcome the main obstacle that DNA entry into the nucleus constitutes, as well as prevent unwanted DNA integration. In addition, delivering the Cas9 protein leads to a better control in Cas9 expression, thereby attenuating off-target Cas9 nuclease activity. This is undoubtedly why scientists need to have a reliable transfection reagent especially designed for this application.

We now provide a full range of reagents for all your CRISPR experiments (Table 1 & Fig. 6):

  • jetCRISPR™ for RNP delivery – Protein and guide RNA co-delivery
  • jetPRIME® for CRISPR DNA plasmid approaches
  • jetMESSENGER™ for RNA transfection (guide RNA and mRNA cotransfection)
  • in vivo-jetPEI® for in vivo gene editing through delivery of DNA (Zuckermann et al. (2015) Nat Commun6, 7391; He et al. (2016) PLoS Pathog 12. E1005743)

 

  in vitroin vivo   

   Delivered molecule   

   Cas9   

   Guide RNA   

   Our solutions   

  in vitro      Protein/RNA    Protein    RNA
   jetCRISPR™
   RNP transfection reagent  
  in vitro      DNA    DNA    DNA
   jetPRIME®
   DNA transfection reagent  
  in vitro      RNA    mRNA    RNA
   jetMESSENGER™
   mRNA transfection reagent  
  in vivo      DNA    DNA    DNA    in vivo-jetPEI®
….in vivo
delivery reagent  

Table 1: Range of transfection reagents for CRISPR experiments.

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

Every batch of jetCRISPR™ reagent is tested in-house in HEK-293 cells following a reverse protocol in 96-well plates with 30 nM of RNP targeting the HPRT1 gene using 0.3 µL of jetCRISPR™. INDEL (%) is quantified 48 hours post-transfection using the T7 endonuclease assay. Each vial is provided with a Certificate of Analysis.