Description
Description Transgenesis platform is intended to produce new genetically modified mouse models by insertional mutagenesis or homologous recombination. The platform allows the establishment of new mouse models by injecting modified ES cells, provided by researchers, into blastocysts. Transgenesis platform is able to conduct all steps leading to the germline transmission of the mutation introduced into the injected cells: blastocysts production, pseudo-pregnant females, micro-injection of ES cells into blastocysts, reimplantation of blastocysts into pseudo-pregnant females, mating of chimeric mouse for germ line transmission.
Contact
Tiffany Marchiol, Engineer, University of Limoges
3C & 4C-seq
CRIBL has been interested for many years in the role of IgH enhancers on CSR and SHM events. New molecular biology techniques and high throughput sequencing allowed to study and decipher enhancers’ study. CRIBL have skills in 3C and 4C-Seq (Circular Chromosme Capture Conformation) techniques, allowing the study of gene interactions at the genome level. The 4C-Seq is carried out in collaboration with Pedro P. Rocha (Unit on the structure and regulation of the genome, NIH).
LAM-HTGTS
Linear amplification-mediated high-throughput genome-wide sequencing (LAM-HTGTS) is a method for genome-wide detection of legitimate rearrangements or translocations from a given location in the genome (called bait).
These 2 coupled techniques make it possible to study the role of enhancers of the heavy chain immunoglobulin locus (Eµ-MAR and 3’RR) on maintaining the organization of the B lymphocyte genome by studying: the spatial organization of the lymphocyte B (ie: folding of the IgH locus on itself and interaction with partner genes) and legitimate (CSR) and illegitimate genomic rearrangements (translocations).
Contact
Sandrine Le Noir, Researcher, CNRS
CSReport
CRIBL has developed bioinformatics tool allowing the study of recombination junctions of the IgH locus (class switch, CSR, and suicide recombination of the locus, LSR): CSReport. This tool was designed to allow automatic analysis from high throughput sequencing data (J Immunol. 2017 May 15; 198 (10): 4148-4155).
CSReport identifies recombination junctions from the alignment of high throughput sequencing reads to reference sequences. CSReport determines the break points on each segment involved in the junction with a resolution of 1 bp and determines the most frequently targeted 8 nucleotide patterns. CSReport analyzes the structure of junctions (junctions with insertion, frank and with microhomologies). Analysis time: ≈ 30 mins/106 séquences.
CSReport requires the Python3 and Jupyter environments (preferably from an Anaconda distribution) with an updated Biopython package. The CSReport notebook file (ZIP archive) including the reference files is available on request.
Contact
Sophie Péron, Researcher, INSERM
DeMinEr
CRIBL has also developed a tool for the study of rare mutations. DeMinEr enables detection of rare AID-induced mutations from high throughput sequencing data from a sequencer-induced error correction approach by sequencing an unmutated sample in parallel (J Immunol. 2018 Aug 1; 201 (3): 950-956).
Contact
Eric Pinaud, Research Director, CNRS
Description
CRIBL focuses its research on both murine and human normal or pathological B lymphocytes. We have developed several routine flow cytometry and fluorescence microscopy protocols to analyse mouse models and human samples.
Contact
Claire Carrion, Engineer, CNRS
Engineering of immunity protein
The CRIBL laboratory has an internal mini-platform for protein production and purification, mainly focused around immunoglobulin. This platform brings together equipment for the overexpression, purification and analysis of proteins.
This platform has access to laboratory facilities: cell culture rooms with hoods, incubators and shakers or a protein analysis room with SDS-PAGE, Western-blot, ChemiDocTouch (Bio-rad) and spectophotometer equipment.
Purification equipment is specific to the mini-technical platform: the protein purification is carried out using an ÄKTA FPLC in a refrigerated chamber with a panel of purification columns (affinity, size exclusion), under the supervision of the manager.
Contact
Christelle Oblet, Engineer, CNRS
Radioimmuntherapy studies
CRIBL has access to a SPECT camera coupled to a scanner (Computerizd Tomography), dedicated to small animals (rat, mouse), allowing imaging after administration of a radiolabelled molecule using a gamma emitter (99mTc, 123I, 111In, etc.).
This technology allows the monitoring of the distribution of this compound in the body.This type of technology can be applied to the study of the biodistribution of a molecule of interest (after development of its radiolabelling in collaboration with the Nuclear Medicine department of the CHU), but also allowing tumoral progression monitoring by CT scan or using tracer available.
Contact
Stéphanie Durand-Panteix, Engineer, University of Limoges
Flow cytometry platform
Flow cytometry platform, localized in the CBRS is a part of the technological platform BISCEm US 42 INSERM / UMS 2015 CNRS.
The flow cytometry platform has the scientific and technical skills for project implementation from sample preparation to data analysis and provides the necessary services and training.Its resources make it possible to characterize and separate populations of cells, both prokaryotes and eukaryotes, or of particles (vesicles, nanoparticles, organelles, etc.) for basic and / or clinical research in immunology and hematology.
The platform offers assistance in the design of experiments (in particular on the choice of fluorochromes) assistance in the analysis, interpretation and presentation of results. It adapts its equipment by responding to the new expectations and needs of users by developing new techniques (flow imaging).
We also offer theoretical and practical training in the use of each analyzer cytometer and, if necessary, the cell sorter. The goal of this training is for all users to quickly become self-sufficient and independent in the use of cytometers
Contact : Catherine Ouk, Engineer, CNRS
Microscopy platform
Microscopy and imaging platform, localized in the CBRS is a part of the technological platform BISCEm US 42 INSERM / UMS 2015 CNRS.
This platform is open to all laboratories at the University of Limoges as well as to external structures (public and private). It provides fluorescence microscopy and image analysis systems for free use or assisted by the platform engineer.
Contact : Claire Carrion, Engineer, CNRS
