Title: Advanced molecular biology tools in dermatological research
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
- 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