Song:Experimental Design For Brainbow
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Aim 1.To generate cell lines random-integrated with brainbow cassette
- To construct lentiviral vector carrying Brainbow-2.1 cassette
- To package and transfect lentivirus
- To isolate GFP-positive cells by flow cytometric cell sorting
- To establish integrated clonal cell line from single cell
- To validate persistent and efficient expression of GFP after passaging and during differentiation
Aim 2.To introduce inducible Cre into brainbow-integrated cell lines
Aim 3.To establish live cell imaging and automated lineage tracking system
Aim 4.To manually isolate single cell from differentiation culture
Aim 5.To perform single-cell transcriptomic and epigenomic analysis