BIOL 2299 Lecture Notes - Lecture 2: Chordin, Amyloid Precursor Protein, Frontal Lobe
1 Jun 2016
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Ways to make ipsc cell: retroviral or lentiviral transduction: genomic integration, but tumors in chimeric animals due to ectopic expression of proviral transgenes. Plasmid transfection: no genomic integration and therefore unlikely to disrupt the host"s genome and cause cancer. Future goal: use small molecules to get these genes into the cell; don"t want these genes constantly expressed, just until you need to differentiate them. Yamanaka: developed the system used to identify the set of genes that could induce pluripotency: 1. Identified 24 genes that are present in embryonic stem cells: 2. Used biotechnology to replace the dna sequence for the fbx15 protein with two genes: one which causes a blue color and another that makes a cell resistant to a drug. Cells that normally express fbx15 would now be resistant to the drug: 3. Engineered retroviruses to express 1 of the 24 genes and transfected cultured mouse fibroblasts (found in the skin) with each virus: 4.
