Overview

Specifications

Reagent

jetCRISPR®

Molecule delivered

Ribonucleoprotein: Guide RNA and Cas protein (Cas9, Cpf1)

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 1250 transfections in 24-well plates or 300 transfections in 6-well plates

Storage

5°C ± 3°C, stable for 6 months (502-01) to at least one year (other packaging sizes) when stored appropriately


Summary

CRISPR/Cas engineered nuclease system is a powerful and highly specific genome editing tool. CRISPR/Cas is a two component system with a guide RNA (gRNA) molecule that drives a Cas nuclease (Cas9, Cpf1) 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 and CRISPR/Cpf1 experiments. jetCRISPR® leads to high CRISPR genome editing efficiency, while maintaining excellent cell viability and morphology post-transfection. Use the leading technology for CRISPR/Cas9 and CRISPR/Cpf1 RNP delivery!

Ordering information

ProductReference NumberProduct size
jetCRISPR®502-010.1 ml
jetCRISPR®502-070.75 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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Description

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 SpCas9 Nuclease 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 (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®).

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 (SpCas9 Nuclease and HPRT1 sgRNA) with 0.3 µl of jetCRISPR® reagent per well of a 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 a 96-well plate. Genome editing was assessed at different time points 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

Videos

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

FAQ

If you have any questions regarding jetCRISPR®, please visit our dedicated Frequently asked questions or contact us.

Applications

CRISPR/Cas9 Experiment

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):

Polyplus-transfection - Table CRISPR reagents

Table 1: Range of transfection reagents for CRISPR/Cas9 experiments.

Fig. 6: Polyplus-transfection - Product range for CRSIPR experiment

Fig 6: Range of in vitro transfection reagents for CRISPR/Cas9 experiments.

CRISPR/Cpf1 Experiment

In addition to Cas9 protein, other Cas RNA-guided endonuclease are available to improve genome coverage where the Cas9 recognition motif is less represented. acidaminococcus Cas12a protein also known as Cpf1 is available to target AT-rich genomic regions. Delivery of Cpf1/gRNA RNP complexes has proven to be more challenging and to this day, only physical methods such as electroporation were recommended by commercial Cpf1 protein providers. With jetCRISPR®, we now offer a fast and reliable transfection method to efficiently deliver Cpf1 RNP complexes in mammalian cells.

Read more…

Quality

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 (SpCas9 Nuclease & gRNA 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.

Testimonials

“Another lab used a comparable protocol with RNAiMAX. jetCRISPR is preferred for its simplicity, lower cost, and higher efficiency.”
 Olivia G., University of Chicago, USA 
  

Protocol

To view our protocols, please fill in the fields below and click download.

Other files

Bibliography

Order by :  
Found 7 results :
Cell Linein vitro
in vivo
Delivered MoleculeReagentResults & Citations
Human primary hepatocytesin vitroRNP (Cas9 Protein + gRNA)jetCRISPR, SpCas9 Nuclease
10-30 % of INDEL at 30 nM RNP
Protocol available
A549in vitroRNP (Cas9 Protein + gRNA)jetCRISPR, SpCas9 Nuclease
40-60 % of INDEL at 30 nM RNP
Protocol available
HEK-293in vitroRNP (Cas9 Protein + gRNA)jetCRISPR, SpCas9 Nuclease
40-60 % of INDEL at 30 nM RNP
Protocol available
HeLain vitroRNP (Cas9 Protein + gRNA)jetCRISPR, SpCas9 Nuclease
40-60 % of INDEL at 30 nM RNP
Protocol available
Caco-2in vitroRNP (Cas9 Protein + gRNA)jetCRISPR, SpCas9 Nuclease
10-30 % of INDEL at 30 nM RNP
Protocol available
Human primary epidermal keratinocytes, Epithelialin vitroRNP (Cas9 Protein + gRNA)jetCRISPR, SpCas9 Nuclease
10-30 % of INDEL at 30 nM RNP
Protocol available
Human primary melanocytesin vitroRNP (Cas9 Protein + gRNA)jetCRISPR, SpCas9 Nuclease
10-30 % of INDEL at 30 nM RNP
Protocol available