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Tebubio's blog - Acting and reacting in life sciences and biotechnologies
  • Home
  • Research areas
    • ADME-Tox
    • Biomarkers
    • Cell Biology and Signalling
    • Cell Sourcing – Cell Culture Technologies
    • Drug Discovery
    • Gene Expression – Molecular Biology
    • Stem Cells
    • Supplying Discovery Tools
  • Contact us
  • Meet the authors
Drug Discovery

Designer Nucleosomes – the next generation of epigenetic substrates!

10/06/2016 by Ali El Baya, PhD No Comments
Histone_Structure_A-01-1

Fig. 1: Mononucleosome structure

In eukaryotic cells DNA is packaged in nucleosome units called mono-nucleosomes which consist of a segment of DNA called core DNA (147 bp in length) wound around a histone octamer (Fig 1.). Histone octamers are assembled from 2 copies of the the core histones H2A, H2B, H3, and H4. These mono-nucleosomes are connected by linker 80bp-long DNA. A fifth histone type of protein, the so-called linker histone H1, binds to the linker DNA close to the entry and exit of the core DNA and is involved in chromatin compaction (Fig 1.).

Substrates for epigenetic enzyme assays and inhibitor screenings

In the past, native or recombinant nucleosomes, single histone proteins, or histone derived peptides have been available for assays with epigenetic enzymes such as histone methyltransferases or histone acetylases and related screenings for inhibitors of the respective enzymes (for an overview about the products available, you might like to read my recent post Find the best epigenetic enzyme substrate for your needs).

modifications and diseases

Table 1: Specific histone modifications can be linked to certain diseases and effector proteins (provided by Epicypher).

As some epigenetic enzymes require highly specific substrates, Epicypher has now launched a new product line – Designer Nucleosomes (dNuc). dNUCs are semi-synthetic nucleosomes incorporating specific histone post-translational modifications. These reagents represent a powerful new technology – critical in understanding chromatin biology and for the development of novel drug targets and precision therapeutics. It is known that highly specific histone modifications can be linked to certain diseases (see Table 1). Table 1 liste the dNUCs which are already available, for further information have a look at our list of Designer Nucleosomes. dNUCs serving as substrates for the most relevant epigenetic enzymes will be added to our catalog in the coming months. If you have specific modifications in mind, which which are know to be optimal for your enzyme of interest, please let me know through the form below. Epicypher might already have this designer nucleosome in their pipeline – or might be able to produce it on a customized basis.

Download the paper here.
To get more insight into the dNuc manufacturing process Epicypher applies, you can download the white paper Not All Designer Nucleosomes are Created Equal: A Tale of Two Cysteines.

The paper compares the two currently used synthetic methods to produce dNUCs, native chemical ligation (NCL) and methyl lysine analog (MLA). The paper shows that NCL – the method used by Epicypher – yields superior nucleosome preparations.

Any questions or comments? Please feel free to contact me with the form below!

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Supplying Discovery Tools

Cost-efficient pre-optimized transfection reagents

by Dimitri Szymczak, PhD No Comments

The main challenge when choosing a transfection reagent is that we don’t know how it will work with our own cell type of interest. It is also time consuming to find the optimal conditions. Well, here’s the solution:  pre-optimised transfection reagents.

eGFP expression revealing transfection efficiency

Pre-optimised transfection of pEGFP-N3 plasmid expressing eGFP under CMV promoter

They already cover more than 39 cell types including MEF, Caco-2, MCF-7, HepG2, Primary Macrophages, Huh-7 and many others.

Sharing our feedback on performances

Using a vector expressing the eGFP (pEGFP-N3) under CMV promotor, we assessed the transfection efficiency. Take a look at the example of results below. They will give you a pretty good idea of what you can expect.

Specific feedback of Genjet transfection

Transfection efficiencies with pre-optimised Genjet reagents

Can we compare the pre-optimised Genjet?

Ok, now you can see we got good feedback. Still, will it be a better solution?

The answer is yes. The comparison with Lipofectamines, Fugene HD and Amaxa reveals that pre-optimised Genjet reagents allow high number of positive cells (dark green below).GenJet-HepG2-Huh7 comparison

You can see on right of each picture just above, that performances are even better with 10% serum, that would be much appreciated by the cells.

Furthermore, this quality is associated to low price. Check it for yourself!

News

Facebook Summer Quizz from June 10th to June 27th 2016!

by Elodie Monin No Comments

This yFacebook Summer Quizear again, many new products have been added to tebu-bio‘s offer. 

Every day, we seek to innovate and meet our customer’s needs… YOUR needs!

Finding new products and new techniques is great, but it’s essential to share them with you. 

It’s time to test your knowledge before your summer holidays…

 

Answer a few easy questions and join in our draw – you might win a Google Chromecast or one of 3 Google Cardboard 3D Virtual Reality Viewers!

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