Today, I’d like to invite you to take a look at a highly efficient and useful kit, which brings together all the required components you need in a complete system for culturing and transfecting human pluripotent stem cells for gene editing.
The PluriQ™ G9™ Gene Editing System includes the G9™ Maintenance Medium and G9™ VTN Recombinant (vitronectin) plate coating for culturing human induced pluripotent (iPS) or embryonic stem (hES) cells in a manner that maximizes transfection by the included EditPro™ Stem Transfection Reagent to transfect genome editing constructs. The EditPro™ Stem is optimized for delivery of CRISPR/Cas9 editing via DNA Cas9 vectors, Cas9 mRNA with DS DNA (G-block), Cas9 mRNA with tracr/guideRNA, or Cas9 protein with tracr/guideRNA. It’s effective for delivery of TALEN and ZFN editing tools as well. With such high efficiency delivery, it’s not necessary to include an antibiotic selection in the editing design.
iPS or hES cells, when grown on the G9™ VTN included in the system in G9™ Maintenance Medium, maintain their pluripotency with a colony morphology which is that of a mono layer and ideal for even, high efficiency transfection when using EditPro™ Stem. The G9™ Versene is used as a gentle, easy, non-enzymatic method of passaging the cells.
EditPro™ Stem, when used to transfect iPS or hES cells as part of the G9™ Gene Editing System, results in the highest efficiency of transfection regardless of what type of substrate is being transfected. As an example, you can see transfection and expression efficiency with a Cas9 DNA plasmid, and indel formation with transfection of mRNA with tracr/guide RNA or Cas9 protein with tracr/guide RNA. All methods give excellent editing results.
To resume, the benefits are…
- High Efficiency Transfection of human pluripotent stem cells delivering DNA, RNA and/or protein
- No need for antibiotic selection
- Low toxicity and does not affect pluripotency
- Easy to use – no special equipment required
- Cost Effective – effective at low amounts of reagent resulting in low cost/well
- Xeno-free, feeder-free culture of human pluripotent cells
- Easy, gentle non-enzymatic passaging compatible with high throughput culture
Some extra reading? Take a look at the protocol..
There is a step-by-step protocol designed by MTI-Globalstem scientists, click here to take a look at it.
What’s in the box…
The components of the PluriQ™ G9™ Gene Editing System (also available separately) are:
- EditPro™ Stem Transfection Reagent, 0.1 ml, optimized for the delivery of DNA, RNA and/or protein into pluripotent stem cells and tested specifically for delivery of CRISPR/Cas9 editing via DNA plasmid, mRNA Cas9 with tracr/guide RNA, or Cas9 protein with tracr/guide RNA. It is effective for TALEN and ZFN editing as well. Available separately as GST-2174 in the 0.25 ml size.
- PluriQ™ G9™ Maintenance Medium Kit (GSK-9001), a defined, xeno-free, feeder-free medium used for expansion and maintenance of human ES and human iPS cells in vitro. Includes PluriQ™ G9™ Basal Medium (500 ml), PluriQ™ G9™ Supplement (50X), Human FGF-2, Human TGF-β1.
- G9™ VTN Human Recombinant, 0.5 µg/ml (GSR-2070, 1 ml) for plate coating. This provides an ideal substrate which leads to the stem cells being more of a monolayer and a bit more spread out which enhances efficiency and evenness of transfection.
- G9™ Versene Solution (1x) (GSM- 1101, 100 ml) for passaging which allows for easy, non-enzymatic, gentle passaging of the cells and is compatible with scale-up to higher throughput cell culture.
Interested in learning more? Leave your questions and comments below, we’ll be pleased to answer!
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