The aim of our technical platform is to provide our expertise and molecular tools to analyze mRNA translation/stability and generate lentiviral constructs.

Our team can provide the following services:

(i) Density gradient centrifugation. Sucrose gradient centrifugation enables the separation of free RNA from ribosome-bound mRNA.

(ii) Polysome isolation/fractionation. The analysis of ribosome-bound mRNA is a powerful approach to investigate translational activity. Our fractionation platform allows the purification of actively translated mRNAs from a wide range of mammalian tissues and cell types, as well as from other organisms (e.g., bacteria, yeast, parasites). Polysomal RNA can subsequently be processed for downstream applications such as RNA sequencing or RT-qPCR analysis.

(iii) RNA stability assays/Ribonucleoprotein immunoprecipitation assays (RIP). Various trans-acting factors, including non-coding RNAs and RNA-binding proteins (RBPs) regulate mRNA fate (translation and degradation) and play key roles in physiological and pathological processes. We provide support in designing and performing RNA stability assays in your in vitro models. In addition, we offer expertise in RIP assays to identify direct mRNA targets of RBPs.

(iv) Molecular cloning, lentiviral constructs. Planning to overexpress or silence a gene of interest in vitro? Or to generate stable cell lines? We offer comprehensive support for cloning strategies and lentiviral production. Our ressources include plasmids for lentiviral packaging as well as constructs for expressing key regulators of mRNA translation and stability (e.g., RBPs, eIF2α).

(v) Hydrodynamic injection of plasmids in mice. For liver-targeted studies, we provide expertise in hydrodynamic tail vein injection to efficiently deliver plasmids to hepatocytes in vivo.

(vi) Analyze, consulting, support and training. We also assist with experimental design, data analysis, and provide tailored training and scientific consulting.

Contact persons :

Dr. Noémie Legrand, Ph.D. (This email address is being protected from spambots. You need JavaScript enabled to view it.) Scientific supervision. +33 (0)3 20 62 68 62.

Pr. Cyril Sobolewski, Ph.D (This email address is being protected from spambots. You need JavaScript enabled to view it.): Academical supervision.

Mrs. Ophélie Bertin, MSc (This email address is being protected from spambots. You need JavaScript enabled to view it.): Technical support.

Références:

Legrand et al, PLoS One, 2014.

Legrand et al, Nucleic Acids Research, 2015.

Legrand et al, Cancers, 2021.

Sobolewski et al, CMGH, 2021.

Sobolewski C, Dubuquoy L and Legrand N, Cancers, 2022.

Sobolewski et al, Cancers, 2022.