Advanced molecular biology tools in dermatological research

 

EADV-ESDR Summer Research Workshop, 4-8 June 2018. Naples, Italy

This workshop was led by Prof Caterina Missero with faculty from ESDR and was specifically aimed at dermatology residents/trainees and/or at PhD students and postdocs affiliated to dermatological research with limited experience in experimental research and in molecular biology.
 
The workshop comprised a mixture of research presentations, interactive and practical sessions. A booklet of methods and protocol in advanced molecular biology was given to participants during the course. Topics covered included:
 
Gene transduction and silencing in epidermal cells:
  • isolation of human and mouse primary keratinocytes
  • keratinocyte culture and keratinocyte/fibroblast co-culture
  • efficient transduction of genes or gene knockdown in keratinocytes
Gene expression in skin in health and disease:
  • RNA isolation from human and mouse primary keratinocytes, from freshly isolated epidermis, and cells of interest from fixed skin
  • quantitative RT-PCR: procedure and data analysis
  • RNA sequencing and data analysis
Genomic editing:
  • designing of gene deletion or gene replacement by CRISPR/Cas9
  • preparation of Cas9-guide RNA vectors
  • assess genome editing efficiency
  • isolation of genome-edited cells