tebu-bio is a European company specialised in providing innovative reagents and laboratory services in Life Sciences
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Gene Editing - CRISPR-Cas9 genome editing reagents

From knock-out to knock-in and everything between

Thanks to CRISPR-CAS9 the gene editing unleash the possibilities for cell line engineering. A complete lost of fonction can be done with knock-out of the target gene or miRNA. Add a tag to a protein, design a SNP, set a point mutation or even more sophisticated modifications are knock-in projects. We also offer the possibility to fine tune the gene expression level thanks to the TUNR technology that is also based on CRISPR-CAS9.

CRISPR-CAS9 principle for Gene editing

Simplifying gene editing

Just mention your cells (cell lines, IPSCs and ESCs) and your target (gene, SNP, miRNA) and get the establish cell line with knock-out in 4 months and with knock-in in 6 months.

The service includes:

  • Design and validation of reagents
  • Monoclonal cell line establishment
  • Genotyping by NGS

The service is done at St Louis (USA) by Canopy's experts.

Do it yourself with a simple box

We offer complete tool box for CRISPR-CAS9 gene editing KO and KI.

  • The gRNA (in orange) is provided as 2 crRNA (custom-designed for your project and in 10 nmol each and tracrRNA (50 µl, 200 µM in TE, pH 7.5)
  • The Cas9 (in blue) endonuclease protein, 62 µM (5 µl, 62 µM in formulation buffer)
  • Donor construct (HR template)—custom-designed for your project in case of knock-in
  • Genotyping primers (forward and reverse primers in 25 nmole each)

 

Would you prefer more sophisticated options?

Alternative CAS9 endonuclease sources

An approach without vectors can be also interesting. It was shown that 5MeC and PseudoU modifications on mRNA improve the translation and reduce the immune response. We provide ready-to-use CAS9 mRNA in 1mg/ml, as affordable catalogue products throughout Europe.

The gene-editing ribonuclear protein (RNP) complex is formed in the cells thanks to optimized translation. It was shown (Hendel et al 2015, Osborn et al 2016) that synthetic and modified sgRNA is highly preferable. The highlighted modification is 2'-O-methyl 3' phosphorothioate (MS) at both extremities of the sgRNA (red star).

Contact us for a quotation.

The transfection can be ensured by a dedicated reagent, CRISPR mRNA-in. And for those who would like to monitor transfection efficiency, there is also an eGFP mRNA similarly modified.

When your work requires several, or recurrent, or even multiplex gene knock-out, a cell line model expressing CAS9 can be helpful. Notably, it is a smart way to explore the effects of loss of function in a pathway and why not make an epistatic study. Such screening is based on the delivery of a dedicated sgRNA libraries into the CAS9-expressing cell line.

  • All-in-one plasmid

One vector expressing the CAS9 and the specific sgRNA with mCherry reporter to monitor the transfection efficiency.

all-in-one vector

Contact us through the form on this page, mentioning your target gene to get a specific plasmid

Donor plasmid for HR and convenient selection

A Donor is required for knock-in and recommended for knock-out to make selection of the modified cells. We offer ready-to-use Donor vectors for each specific project. The vectors are also available in a cloning version.

 

Contact us through the form on this page to get advice on the best donor plasmid for your project

Adenovirus solution for hard-to-transfect cells

Adenovirus is a means to efficiently deliver the CRISPR-CAS9 system into cells, even for hard-to-transfect cells. Furthermore, adenovirus provide transient expression and no integration in the genome. It's very well adapted to the CRISPR-CAS8 based genome editing.

We offer control adenovirus to monitor the efficiency of the delivery. Also, tailored adenovirus productions including the all-in-one and the donor vector.

 

 

Useful accessory tools

T7 endo nuclease

The generated insertions and deletions can be revealed with a simple T7-Endonuclease I assay.

PolyJet, transfection reagent for large plasmid

CRISPR-CAS9 often involve large plasmid that can be challenging to deliver into cells. We recommend the PolyJet for several reasons.

  •  Bio-degradable after endocytosis - no toxicity
  • Efficient even with very long DNAs (>89 kb)
  • Equally good for both single DNA transfection and multi DNA co-transfection
  • Validated with 80+ cell lines
  • Simple & robust transfection procedure
  • Very affordable

IVT for sgRNA

Despite sgRNA produced by in vitro transcription are not being the most efficient ones, they are functional in numerous articles. To help you with this, we provide a fast and robust in vitro transcription kit called T7-FlashScribe.

FISH probes for chromosome enumeration and ploidy determination

Thanks to our VividFISH Chromosome FISH Probes, control the number of chromosomes and so alleles for specific site.