Biochemistry and Molecular Biology Facility

BIOMOL / BIOCHEM

Biochemistry and Molecular Biology

The Biochemistry and Molecular Biology platform was created in 2012.

Any iBV member who do not have much knowledge and experience in this area can take advantage of the expertise given by people dedicated to the platform (1 engineer).

Platform allows :
1. accessibility to common ressources,
2. skilled personnel to help in experiments.

1. Molecular Biology platform

Presentation

Molecular Biology platform performs constructions for CRISPR/Cas9.
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 model organisms.

The goal of the platform is to make CRISPR/Cas9 constructions and provide validated ready-to-express constructs for any iBV members.

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.

Source : Feng Zhang/MIT

Nowadays, the CRISPR-Cas9 system is a widely used gene editing tool and a platform dedicated to this technique has been created in 2012.

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.
  • Testing of guide RNA design efficiency by in vitro cleavage assay of specific PCR products using in-house produced cas9/sgRNA RNP.
  • Cloning of CRISPR components in a suitable delivery system (plasmid, lentiviral vector, mRNA) and sequence validation.

 

2. Biochemistry platform

Presentation

The platform is dedicated to the expression and purification of proteins, from expressing cell/bacteria preparation to the pure protein.

Techniques and expertise

The strong need for the production of recombinant proteins called for the reinforcement of this activity as a central resource and expertise. The Biochemistry platform provides purified recombinant proteins needed for biochemical screening, as growth factors for cell biology as well as antigens for antibody production using bacterial expression systems.

The facility also houses expertise and equipment to perform:

  • Extraction and renaturation in vitro of bacterial inclusion bodies,
  • Exclusion chromatography (gel filtration / desalting),
  • Ion exchange chromatography,
  • Affinity chromatography (Antibody purification) using tags (His, GST, etc),
  • Hydrophobic interaction chromatography (HIC)/Reverse Phase (peptide purification).
  • An aliquot of purified product is analyzed on SDS-PAGE and, if necessary by Western blotting.

FPLC (Bio-Rad – NGC)

FPLC (Pharmacia – AKTA pure)

Imager (VILBER LOURMAT - FUSION FX7)

Imager (ODYSSEY – LiCor)

Microvolume Spectrophotometer (DeNovix – DS11)

Vaccum Concentrator (EPPENDORF - Concentrator Plus)

Lyophilisator (HETOSICC – Flexi Dry)

1. Molecular Biology platform

Engineer Responsible

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

Scientific coordinators

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

2. Biochemistry platform

Engineer Responsible

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

Technical staff

Maurice Hattab
Tel : +33 4 92 07 64 47
E-mail : hattab@unice.fr

Scientific Coordinators

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

iBV - Institut de Biologie Valrose

"Sciences Naturelles"

Université Nice Sophia Antipolis
Faculté des Sciences
Parc Valrose
06108 Nice cedex 2