CRISPR genome editing: which cell line to choose?

Many labs have adopted the CRISPR genome editing technology to make knock-out and knock-in cell lines.

This technology produces first a targeted break in genomic DNA, which can then be exploited to produce cell lines with genes knocked out or where a donor vector has been used to introduce new genetic elements (point mutants, fluorescent tags, antibiotic resistance cassettes, etc.). Essentially any desired modification to the cells genome can be made. In setting up these genome editing projects there are many choices to be made including vector for the Cas9 protein and for the sgRNAs. Perhaps the most difficult choice, however, can be which cell line to use. Even the most affordable stable genome editing cell line development services can come with a significant cost, so choosing the right cell line at the beginning is crucial. Here we explain some of the choices researchers have in setting up their CRISPR genome editing projects and give our advice for cell line selection.

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Cellular models for studying the human urogenital system

The urogenital system is the organ system of the reproductive organs and the urinary system. These are grouped together because of their proximity to each other, their common embryological origin and the use of common pathways, like the male urethra.

Today, I’ll be taking a look at human primary cells derived from urogenital organs (and in one of my next posts, we’ll explore models for the digestive system.

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Hepatic Cellular Models

A wide range of in vitro models are used in preclinical drug testing for the investigation of ADME-Tox (Absorption, Distribution, Metabolism, Excretion and oxicity) properties of New Chemical Entities (NCEs). The liver is the main organ with regards to ADME-Tox, it’s the place of more than 500 different functions, including: metabolism of lipids, carbohydrates, and vitamins, detoxification, production of bile, albumin, fibrinogen, globulin, etc (1). The liver lobule is composed of parenchymal cells (hepatocytes) and nonparenchymal cells (Kupffer cells, hepatic stellate cells, and sinusoidal endothelial cells).

Hepatocytes – the Gold Standard

Hepatocytes represent 80% of liver volume. Hepatocytes are commonly used in drug discovery and preclinical drug development to perform experiments requiring intact cellular systems. Intact hepatocytes contain the major hepatic drug-metabolizing enzymes required to study the four categories of xenobiotic biotransformation: hydrolysis, reduction, oxidation and conjugation.

Because of these enzymes, hepatocytes provide a viable and cost-effective alternative to in vivo testing. [Read more…]

Zebrafish – an ideal model for morphogenesis analysis

Cdh1 antibody - Rabbit Polyclonal antibody to E-cadherin (Gentex | tebu-bio)

E-Cadherin (Cdh1) expression in Zebrafish with Genetex antibody (GTX125890 | tebu-bio)

In a recent study, Xiong F. et al. have developed a robust Zebrafish-based model allowing the analysis of the impact of cell shape and division orientation on epithelial development. This model explores possible links between cell shape and division orientation in epithelial diversity.

Among all the research tools used, the team of scientists investigated Cdh1 expression by IHC-Wm with a Genetex antibody, dedicated to Zebrafish studies (cat. nr GTX125890). This unique antibody is available in Europe thorugh tebu-bio for IHC applications (recommended dilutions 1:100-1:1000) including IHC Whole mount (IHC-Wm).

This emphasizes the need for researchers to have access to high quality and validated antibodies focused on Zebrafish model, as described in a previous post, showing the benefits of using such primary antibodies in Zebrafish whole mount IHC staining.
Source:
Xiong F. et al. “Interplay of Cell Shape and Division Orientation Promotes Robust Morphogenesis of Developing Epithelia” Cell Volume 159, Issue 2, p415–427. DOI: http://dx.doi.org/10.1016/j.cell.2014.09.007

Human Primary Sebocyte assay for compound testing on lipid metabolism

Sebocytes belong to the epidermal ecosystem together with hair follicules or in sebaceous glands. Sebocytes play a key role in skin health by producing sebum. These cells are now at the center of researchers’ attention for skin research, cosmetology and dermal discovery.

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5 reliable research tools to study ERG, USP9X and DUB inhibitor in prostate cancer

In a recent paper published in PNAS, Wang et al. show that the inhibition of the deubiquitinase USP9x by the small molecule WP-1130 is able to induce degradation of the transcription factor E-twenty-six related gene (ERG).

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