Functional genomics
Team :
Sandrine Baghdoyan : Research Engineer INSERM
Delphine Laustriat : PhD Student
Jacqueline Gide : Associate engineer (CECS)
Laetitia Barrault : Associate engineer (CECS)
Ting Ting Qing: Post-doctoral Fellow
The main topic of our team is the development of biotechnological tools for the study of monogenic diseases. Our activities are subdivided into 3 projects:
● Functional genomic screens dedicated to monogenic diseases by genes overexpression or extinction
● Differential microRNA profiling
● Inducible pathological hES cell lines
Currently, the proof of concept of all these approaches is carried out on the Myotonic Dystrophy type 1 (DM1) also called Steinert disease.
● Functional genomic screens dedicated to monogenic diseases by genes overexpression or extinction
● Differential microRNA profiling
● Inducible pathological hES cell lines
Currently, the proof of concept of all these approaches is carried out on the Myotonic Dystrophy type 1 (DM1) also called Steinert disease.
Functional genomic screens dedicated to Steinert disease
We are working on human embryonic stem cell lines carrying the causal mutation of DM1. These mutated cell lines were characterized in the laboratory by the “Neuromuscular diseases” I-Stem’s team, led by Dr Geneviève Piétu, which valids the expression of structural and functional phenotypes characteristic of the pathology, both in undifferentiated hES cells and in their progenies.
We want to identify new genes able to disrupt mutant phenotype expression in a mesodermic progeny (MPC) from a DM1 hES cell line. To this aim, we develop screenings of siRNA collections (small interfering RNA) (PARI Platform collaboration) and plasmidic expression vectors (Genoscope collaboration) by high throughput transfections in 96 and 384-well plates, in order to test the effect of the extinction or the overexpression of human genes on DM1 mutant phenotype.
We want to identify new genes able to disrupt mutant phenotype expression in a mesodermic progeny (MPC) from a DM1 hES cell line. To this aim, we develop screenings of siRNA collections (small interfering RNA) (PARI Platform collaboration) and plasmidic expression vectors (Genoscope collaboration) by high throughput transfections in 96 and 384-well plates, in order to test the effect of the extinction or the overexpression of human genes on DM1 mutant phenotype.
Screening of microRNA associated to Steinert disease
In collaboration with Xavier Nissan from the I-Stem’s qPCR platform, we compare expression profiles of 365 microRNA extracted from a DM1 hES cell line to those extracted from a native hES cell line. We identified one group of microRNA strongly induced in DM1 hES and in several tissues from DM1 patients.
Functional assays will be done in order to explore the relation between the expression of these microRNA and the expression of DM1 cellular and molecular phenotype in hES and their progeny.
Functional assays will be done in order to explore the relation between the expression of these microRNA and the expression of DM1 cellular and molecular phenotype in hES and their progeny.
Development of an inducible DM1 hES model for the study of Steinert disease
The screening of siRNA, cDNA and microRNA will allow us to identify molecular targets whose implication in DM1 should be confirmed.
To this aim, we develop a genetically modified hES cell line by the introduction of a construction allowing the controlled expression of a minigene able to induce DM1 associated cellular damages.
This construction, given by Pr Tom Cooper, Baylor College of Medecine, Houston, Texas, USA, was validated in vivo in a model of DM1 transgenic mouse (Orengo, JP., PNAS, 2008).
Such a DM1-induced cellular model will allow us to compare the impact of one gene overexpression or extinction in native or mutant cells populations sharing the same genetic background, except by the expression of the mutant gene.
To this aim, we develop a genetically modified hES cell line by the introduction of a construction allowing the controlled expression of a minigene able to induce DM1 associated cellular damages.
This construction, given by Pr Tom Cooper, Baylor College of Medecine, Houston, Texas, USA, was validated in vivo in a model of DM1 transgenic mouse (Orengo, JP., PNAS, 2008).
Such a DM1-induced cellular model will allow us to compare the impact of one gene overexpression or extinction in native or mutant cells populations sharing the same genetic background, except by the expression of the mutant gene.
