Nov

25

- (1) Targeted Genome Editing using CompoZr ZFN and CRISPR Technology - (2) Duolink Reagents for Localization and Quantitation of Protein-Protein Interactions

Tuesday, November 25, 2014, from 1:45 PM to 4:00 PM

- Targeted Genome Editing using CompoZr ZFN and CRISPR Technology

The discovery of the type II prokaryotic CRISPR/Cas immune system has allowed for the development of an RNA guided genome editing tool that is simple, easy and quick to implement. Sigma Aldrich have developed a single vector system with a Cas9-GFP and a U6-guideRNA expression cassette for CRISPR/Cas9 delivery and expression. CRISPR/Cas systems along with our CompoZR® ZFN technology provide a quick and affordable solution to targeted genome editing in a wide variety of species and cell types.
This presentation highlights how both technologies can be utilised to generate precisely targeted genome edits. We will also discuss the use of high fidelity paired nickases as a means of increasing CRISPR specificity. (45 mn)


- Duolink Reagents for Localization and Quantitation of Protein-Protein Interactions

Duolink reagents are based on in situ proximity ligation assay (PLA) technology, which enables researchers to detect and individually count single endogenous proteins, protein-protein interactions and their modifications in fixed cells and tissues with a fast and simple protocol.

Duolink allows the user to combine any pair of immunofluorescence or immunohistochemistry validated antibodies to fit their unique application, and the read-out is performed either with a fluorescent probe, for fluorescence microscopy, or HRP for brightfield detection. The resulting distinct spots are derived from single-molecule events which are visualized using a standard microscope. Single proteins, protein-protein interactions (including homodimerization events), and protein post-translational modifications are visualized in fixed cells and tissue samples with very high specificity in their correct cellular context at physiologically relevant expression levels even for transient events. Hence, the Duolink assay is a very advantageous choice for unraveling important biological events when compared to traditionally used technologies such as Co-IP. (40 mn)

Lecturer

SIGMA ALDRICH

Contact

Alain Buisseret
SIGMA ALDRICH

Location

de Visscher Auditorium
Avenue Hippocrate 74 (Ground Floor)
1200 Brussels
Belgium