Get up to 85% cleavage efficiency by pairing GeneArt Platinum Cas9 Nuclease and Lipofectamine CRISPRMAX Cas9 Transfection Reagent

Invitrogen™ GeneArt™ Platinum Cas9 Nuclease is wild type Cas9 in protein form for genome editing with CRISPR-Cas9 technology. Cas9 protein and guide RNA (gRNA) form a very stable ribonucleoprotein (RNP) complex that can act immediately after it enters the cell, since transcription and translation are not required. Moreover, the complex is rapidly cleared from the cell, minimizing the chance for off-target cleavage events when compared to vector-based systems. 

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Award-winning Platinum Cas9 Nuclease

“The things that we knew were important were consistent quality, activity, and purity,” says Jason Potter, Thermo Fisher senior staff scientist and R&D manager of genome-editing tools and synthetic biology. “So we made sure, through extensive testing, that we had a very robust purification process.”

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The Cas9 RNP complex can act immediately after it enters the cell, since transcription and translation are not required. Moreover, the complex is rapidly cleared from the cell, minimizing the chance for off-target cleavage events when compared to vector-based systems. 

Cas9 RNP

Review validation data in multiple cell types

Amazing transfection efficiency in over 20 cell lines
Lipofectamine CRISPRMAX reagent effectively delivers CRISPR-Cas9 protein complexes, resulting in cleavage efficiencies on par with electroporation. The table below summarizes cleavage efficiencies as measured by the % indel (insertions and deletions) for lipid-mediated delivery or electroporation, demonstrating successful genome editing in standard and difficult-to-transfect cell lines. Cleavage efficiency has been demonstrated in over 20 cell types, including iPSCs, mESCs, N2A, CHO, A549, HCT116, HeLa, HEK293, and several others.

Comparison of lipid-mediated transfection and electroporation delivery and resulting cleavage efficiencies (% indel) for 20 cell lines.

    % Indel
Cell type Cell type source
 Lipofectamine
CRISPRMAX Transfection Reagent		 Electroporation
HCT116 Human colon carcinoma 85 ± 5 —
293FT Human kidney 85 ± 5 88 ± 3
mESC Mouse embryonic stem cell 75 ± 3 74 ± 4
HEK293 Human kidney 75 ± 3 —
N2A Mouse neuroblastoma 70 ± 5 81 ± 2
3T3 Mouse embryonic fibroblast 57 ± 4 50 ± 2
CHO-K1 Chinese hamster ovary 57 ± 1 —
iPSC Human episomal induced pluripotent 55 ± 3 85 ± 2
U2OS Human osteosarcoma 55 ± 4 70 ± 3
HeLa Human cervical cancer 50 ± 7 —
A549 Human lung carcinoma 48 ± 3 66 ± 3
COS-7 Monkey kidney
 44 ± 3 —
MDA-MB-231 Human breast cancer 39 ± 5 —
HepG2 Human liver cancer 30 ± 3 52 ± 3
K562 Human lymphoblastoid 20 ± 2 91 ± 1
Jurkat Human T leukemia 19 ± 3 94 ± 2
HEKa Human primary epidermal keratinocytes 14 ± 2 32 ± 2
THP-1 Human monocytes 12 ± 3 31 ± 3
HUVEC Human umbilical vein endothelium 9 ± 3 26 ± 2
MCF-7 Human mammary gland 8 ± 4 22 ± 5
(—) not tested

Observe high cleavage efficiencies

Fewer cells needed to initiate your experiment

Lipofectamine CRISPRMAX reagent is extremely gentle on cells compared to electroporation and other reagents. Cells have a high survival rate 48 hours after transfection (panel A), reducing the number of cells you need from start to finish.

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Panels A and B. The cell morphology images captured from an Thermo Scientific™ EVOS™ XL imaging system and the viability graphs provide evidence of Lipofectamine CRISPRMAX’s low toxicity, even after 48 hours of incubation.

Preformed Cas9 protein–gRNA RNPs

Our R&D scientists demonstrated that by combining CRISPR-Cas9 protein with predesigned, pre-transcribed (in vitro) gRNAs and complexing them with a lipid-based transfection reagent, you can significantly simplify the cell engineering workflow and achieve cleavage efficiencies of up to 85% in many cell lines ( Liang et al. 2015).

Lipofectamine CRISPRMAX transfection reagent represents just the vehicle for the Cas9 RNP complex. In addition, the Cas9 protein contains 3 nuclear localization signals to promote efficient transport into the nucleus whether you choose transfection or electroporation for delivery.

Transfection-ready format with >99% purity minimizes reagent preparation and simplifies high-throughput and multiplex genome editing. The Cas9 nuclease:

Enhance protein complex delivery

With the new Invitrogen™ Lipofectamine™ CRISPRMAX™ transfection reagent, it is now possible to use a lipid-based reagent to deliver these protein complexes. Lipofectamine CRISPRMAX reagent is the first optimized lipid nanoparticle transfection reagent for CRISPR-Cas9 protein delivery, providing up to 85% cleavage efficiency when combined with GeneArt Platinum Cas9 Nuclease. In addition, Lipofectamine CRISPRMAX reagent is gentle on cells and cost-effective.

Get our superior Invitrogen™ GeneArt™ Platinum™ Cas9 Nuclease as well as other CRISPR-Cas9 proteins with a reagent that provides:

  • Demonstrated cleavage efficiency in over 20 cell types, including iPSCs, mESCs, N2A, CHO, A549, HCT116, HeLa, HEK293, and several others (See Indel Efficiencies tab on this page)
  • Gentle transfection (low toxicity), meaning fewer cells needed to initiate your experiment  (See Indel Efficiencies tab on this page)
  • The lowest reagent cost option, whether cost per reaction or initial investment
  • An ideal delivery solution for high-throughput experiments

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