Lentivirus production and infection were performed as previously described21 (link). Briefly, HEK293T cells were transfected with third generation packaging plasmids and individual sgRNA vectors or CDKO libraries. Lentivirus was harvested after 48 h and 72 h and filtered through a 0.45 μm PVDF filter (Millipore). Spin infections were used to deliver sgRNA vectors and libraries into K562 cells.
Protocol detail
Lentiviral Transduction of Cells
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Other organizations : Stanford University
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Variable analysis
independent variables
- Third generation packaging plasmids
- Individual sgRNA vectors
- CDKO libraries
- Lentivirus production
- Lentivirus infection
- HEK293T cells
- K562 cells
- Incubation time (48 h and 72 h)
- Filtration through 0.45 μm PVDF filter
- Spin infections
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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