Researcher engineer in Gene therapy for a severe genetic skin disease using conjugated siRNAs

Description de l’employeur

The candidate will join an international research laboratory of 18 persons with a strong and long lasting expertise in genetic skin disorders, gene therapy, tissue engineering and in murine models of genetic skin diseases. He/she will have a unique opportunity to be trained in cutting edge technics in keratinocyte biology and gene replacement therapy. The laboratory is part of the Imagine Institute for genetic diseases located at Necker hospital for sick children. The Institute has high-performance technical platforms in genomics, bioinformatics, cell imaging, flow cell sorting and animal models which could assist the applicant if necessary. The Institute gathers research teams with complementary expertises in multiple aspects of genetics, immunology, cell biology and bioinformatics. The Institute is highly dynamic and will allow multiple interactions with leading scientists in gene therapy, immunology and human genetics (http://institutimagine.org).

Missions et activités

He/she will use a bicystronic expression system enabling the screening of the candidate siRNAs molecues in HEK cells. This system will comprise a fluorescent marker downstream of the wild-type or mutant specific keratin cDNA as peviously used in our laboratory (Pendaries et al . JID 2022). This expression system will be used to screen the efficacy and specificity of conjugated siRNA molecules. The siRNA molecules allowing the best differential inhibition of mutated keratin transcripts will be transfected in patient primary keratinocytes carrying each of the selected frequent keratin mutations. Efficacy will be assessed by quantifying wild-type and mutated keratin transcripts using allele specific qRT-PCR and droplet digital PCR. Spontaneous or induced keratin aggregates will be analyzed by confocal microscopy and/or electron microscopy.