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.
The candidate will :
- Execute protocols
- Perform tissue/molecular analyses of biological samples (immunofluorescence staining, microscopy, RT-qPCR, etc.)
- Manage the order, storage and annotation of biological samples
- Keep an impeccable record of experimental data/measurements
- Perform bibliography search and stay up-to-date with the technical advances in the field
- Interact with a very dynamic scientific team
Fourchette de salaire : (choisir la catégorie incluant 38 400 euros brut par an) Qualifications :
Candidates should have a BTS/DUT, or License degree with a strong background in cell biology and molecular biology.
- We are looking for a highly motivated scientist with a strong experience in molecular cloning and cellular biology.
- A previous experience in gene therapy would be an advantage.
- Good communication skills in oral and written English
- Excellent organization skills
- Ability to work in a team
The candidate should have been able to identify the most effective and specific conjugated siRNAs for 2 frequent keratin mutations and have assessed whether they reverse the disease phenotype (keratin aggregation) in a 2-D culture system using patient primary keratinocytes
How to apply
Please contact Alain HOVNANIAN including in the message a letter of motivation and a CV.