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A Protocol for the Production of Integrase-deficient Lentiviral Vectors for CRISPR/Cas9-mediated Gene Knockout in Dividing Cells
Capitoli
- 00:05Titolo
- 00:52Transfecting of HEK-293T Cells Using Calcium Phosphate
- 02:19Harvesting Virus
- 02:50Concentration of Viral Particles by Ultracentrifugation
- 05:11Esimating of Viral Titers Using p24-enzyme-linked Immunosorbent Assay
- 07:31Counting GFP-positive Cells
- 08:25Results: Production and Validation of the Knockout-efficiency of Integrase-deficient Lentiviral-CRSPER/Cas9 Vector
- 09:57Conclusion
We describe the production strategy of integrase-deficient lentiviral vectors (IDLVs) as vehicles for delivering CRISPR/Cas9 to cells. With an ability to mediate quick and robust gene editing in cells, IDLVs present a safer and equally effective vector platform for gene delivery compared to integrase-competent vectors.
