KEY WORDS
DNA methylation, epigenetics, CRISPR, digestive system, RNA transfection
WORKING ENVIRONMENT AND ACTIVITIES
You will be based in the "Genetics, Physiology and Farming Systems" joint research unit, in a team
working on the genetic and epigenetic determinism of traits in farm animal species.
The aim of this post-doctorate is to develop epigenome editing (notably DNA methylation) in
cellular models of the pig digestive epithelium in order to repress the expression of target genes of interest
for intestinal barrier function. These epigenomic editing tools could then be used in the future for
functional genomics studies on cellular models, in order to limit the need for animal experimentation.
Epigenome editing tools are mainly available in the form of plasmids for DNA transfections. They
are based on the targeting by a guide RNA of an inactivated Cas9 (dCas9) coupled to one or more
epigenetic enzymes (e.g. methyltransferases). Our cell models require high transfection efficiencies, which
are more easily achieved by transfection of messenger RNA. You will therefore adapt and optimise existing
plasmids for in vitro transcription, while studying the efficiency and specificity of epigenome editing
inductions. Among the possible optimisations you will explore are:
- the use of dCasMini as a shorter alternative to dCas9
- the use of the bacterial methyl-transferase MQ1 as an alternative to DNMT3A/L
- direct coupling between the guidance system and the epi-editing enzyme(s), or use of a SunTag
secondary coupling
Your immediate working environment includes experts in DNA methylation, cell biology and the
physiology of the digestive system. You will be able to take part in the activities of the INRAE 'genome
editing' and 'organoids' working groups, as well as in the activities of the European GEroNIMO consortium
(www.geronimo-h2020.eu). You will benefit from premises, platforms and technical support adapted to
your work.
More specifically, you will :
- be responsible for the design, sub-cloning and evaluation of epigenome editing tools. You will
share the tools developed with the scientific community via the addgene plasmid sharing
platform. - study the specificity, longevity and impact on transcription of epi-editions on candidate genomic
regions. You will carry out functional tests of the barrier function of the intestinal epithelium
using cell models of the porcine intestinal epithelium (immortalised cell lines). - disseminate your results in the form of presentations within national and international networks
and scientific articles. - take part in writing a prospective report on epigenome editing in farmed species, covering
technical, scientific, ethical and economic aspects, in collaboration with experts on the subject
as part of the European H2020 GEroNIMO project.
Special conditions of activity: This post-doctorate can be extended, in particular to develop epigenome
editing on intestinal organoids, explore functional impacts in relation to pathogen resistance, or explore the
feasibility of oral transfection of the digestive tract in vivo.
TRAINING AND SKILLS REQUIRED
- Minimum degree required: PhD
- Recommended training: Molecular and cellular biology
- Knowledge required: Transcriptional regulation, epigenetics, genome editing
- Appreciated experience: Subcloning, eukaryotic cell culture, fluorescent microscopy, nucleic acid
extractions, PCR - Skills sought: Scientific writing and oral presentation in English
CONDITIONS
Please do not hesitate to contact Guillaume Devailly (guillaume.devailly@inrae.fr) for further information,
and to apply if you are interested in the subject without mastering all the technical prerequisites.
24 months to start in january 2024
Gross salary 2643€ per month