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Dermatology
Molecular Biology
Built from Dermatology, 5th Edition
What’s inside
7 sections · 70 slides
Overview
- What this topic covers
- Why dermatologists need molecular biology
Tissue and Cell Preparation
Getting a biopsy ready for DNA, RNA, or protein study
- Tissue processing options
- Routes for processing a tissue sample
- Laser capture microdissection
- Laser capture microdissection
- Flow cytometry and cell sorting
- How flow cytometry and FACS work
Core Techniques for DNA, RNA, and Protein
Extract the molecule of interest, amplify it, then detect it
- The shared logic behind molecular methods
- Polymerase chain reaction (PCR)
- The PCR cycle
- Reading the whole genome fast
- Next-generation sequencing workflow
- Sanger DNA sequencing
- Sanger sequencing detection
- Reverse transcription PCR (RT-PCR)
- How RT-PCR converts RNA to DNA
- Western blot (immunoblot)
- Western blot technique
- ELISA and multiplex bead assays
Measuring Cells Within Tissue Sections
Seeing where a gene or protein sits inside intact tissue
- Fluorescence in situ hybridization (FISH)
- How FISH detects gene abnormalities
- Immunohistochemistry
Measuring the Transcriptome and Proteome
Reading nearly all genes or all proteins in one experiment
- Measuring the transcriptome
- Nucleic acid array formats
- Measuring the proteome
- Mass spectrometry proteomics workflow
Mouse Models of Skin Disease
Testing what a gene does by adding or removing it in a living mouse
- Transgenic mice
- Making a transgenic mouse
- Knockout transgenic mice
- Making a knockout mouse
- Conditional knockout mice
- Cre-lox conditional knockout system
- CRISPR-Cas9 genome editing
- Two outcomes after a CRISPR cut
- CRISPR-Cas9 mechanism
Gene-Based Therapy for Skin Diseases
Using DNA and RNA tools to treat disease at its genetic source
- Principles of gene-based therapy
- In vivo versus ex vivo delivery
- Stem cells and induced pluripotent stem (iPS) cells
- Delivering a replacement gene copy
- Correcting the endogenous gene
- Silencing mRNA with siRNA and miRNA
- Gene silencing via mRNA knockdown
- Ex vivo skin gene therapy
- Polymerase chain reaction
- Polymerase chain reaction (continued)
- Massively parallel sequencing
- Massively parallel sequencing (continued)
- Reverse transcription PCR
- Reverse transcription PCR (continued)
- Western blot
- Western blot (continued)
- Fluorescence in situ hybridization (FISH)
- Fluorescence in situ hybridization (FISH) (continued)
- Proteomics with mass spectrometry
- Proteomics with mass spectrometry (continued)
- Transgenic mice
- Transgenic mice (continued)
- Knockout transgenic mice
- Knockout transgenic mice (continued)
- Skin gene-based therapy
- Skin gene-based therapy (continued)
- 30-40
- Where the field is heading
- References
- References (continued)
- References (continued)
- References (continued)
- References (continued)
- Dermatology, 5th Edition (2-Volume Set)