Description of the Molecular Biology Platform

 

The Molecular Biology platform develops innovative approaches using the CRISPR/Cas9 technology which is now recognized as the most efficient gene editing method. The mission of the platform is to provide access to the latest technologies available for introducing targeted genome alterations (delete, edit, or insert genes ) in various life model organisms and in cultured cells.

The goal of the platform is to test, develop and apply modified CRISPR/Cas9 protocols and provide validated ready-to-express constructs.

 

Platform general functioning

In order to develop and optimize new protocols related to CRISPR/Cas9, the Molecular Biology Platform of the Institute of Biology-Valrose (iBV) was created in 2013, with a full-time engineer. This facility, is located on the 2nd floor of the Sciences Naturelles building of the Valrose campus. The know-how of this facility is provided in priority to any iBV member who do not have much knowledge and experience in this area and can take advantage of the expertise given by the permanent engineer dedicated to the platform. The use of the platform is made through an online reservation system in the intranet iBV website. The platform is also open to external academic users upon reservation with adapted rules and special fees. The consolidation of this platform by the present demand is essential to meet the growing demand of the scientific community, and to explore many possible applications. In this context, our platform, dedicated to the modification of the genome, will be extremely attractive and may serve as a regional reference for other institutes in the PACA region.

 

Techniques and expertise

The CRISPR/Cas is an adaptative bacterial defense mechanism that serves to recognize and cleave incoming foreign nucleic acids. This system has recently been adapted to perform genome modifications in other organisms including humans (Cong et al., 2013; Mali et al., 2013), mice (Wang et al., 2013), zebrafish (Hwang et al., 2013) and Drosophila, C. elegans , sea urchin and others.

 

 

   Source : Feng Zhang/MIT

 

Services

The platform offers a full line of CRISPR-Cas9 services to researchers, as well as help for designing and implementing projects.

All projects include:

  • Designing optimal CRISPR-Cas9 target sites for your model organism and PCR primers for the target with the help of the following web sites:
    http://www.e-crisp.org/E-CRISP/
    http://crispr.mit.edu/
    http://tools.flycrispr.molbio.wisc.edu/targetFinder/
    http://www.flyrnai.org/crispr/
     
  • Testing the efficiency of our guide RNA design by in vitro cleavage assay of specific PCR products using in-house produced Cas9 protein and sgRNA.
  • Testing delivery system (plasmid, lentiviral vector, mRNA)
  • Cloning of CRISPR components in delivery system and sequence validation.
  • Researchers validate targets and efficiency in transgenic animals or in somatic (mitotic and post-mitotic) tissues or in cultured cells.

 

 

See annex for more details on various projects

 

 

 

 

 

CONTACTS
Engineer Responsible :

Virginie Virolle
Tel : +33 4 92 07 68 02
E-mail : vvirolle@unice.fr

Scientific committee :

Pascal Thérond
Tel : +33 4 92 07 64 46
E-mail : therond@unice.fr

 

Jean-Claude Chambard
Tel : +33 4 93 37 77 07
E-mail : chambard@unice.fr

 

Laurent Ruel
Tel : +33 4 92 07 64 58
E-mail : ruel@unice.fr

 

 
Molecular Biology Platform
--
Institut de Biologie Valrose
--
CNRS UMR7277
Inserm U1091
--
Université Nice Sophia Antipolis
--
Parc Valrose
06108 Nice cedex 2
France
 

 

 
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CNRS UMR7277, Inserm U1091, UNS Université Nice Sophia Antipolis
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