Epigenetic Programming L8

Epigenetic Programming L8 - Epigenetic Programming (Part...

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Epigenetic Programming (Part II) - Remember that lentivirus and retroviruses are used for genomic integration - RNA can be microRNA or mRNA - Know the four transcription factors (OSKM) in several approaches to generate iPS cells - The retrovirus is used to carry 4 transcription factor genes or mRNA into fibroblast cells o Get vector to carry the gene of interest Ex. Four transcription factor genes o Fibroblast cells then express mRNA and protein - Can also directly insert protein into cell- process inefficient o Transporter (channel) used o HIV virus can also be used to transport proteins - Also rembember DNA and RNA transport (first column) Adenovirus Vector - Day 2- Add adenovirus - 4 transcription factor genes can be inserted into virus - Transduction- add chemicals to enduce virus to get into host cells - Drug selection- getting rid of non-transfected cells - Only cells infected with virus will survive - After cell proliferation of fibroblast cells, adenovirsus will not replicated o Must be re-administered Using Sendai Virus Vector - Don’t need to insert their genes into host DNA - Most of their work is done in the cytoplasm - Used for clinical studies Episomal Vectors - Derived from Epstein-Barr Virus - Plasmid-based genetic reprogramming o Target genes (Oct3/4, Nanog, etc. .) are integrated into plasmid o Heating fibroblast cells will allow reprogramming (15 days are needed)
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Epigenetic Programming L8 - Epigenetic Programming (Part...

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