Category: Cellular Technologies
The CRISPR/Cas9 gene editing system is a very popular tool for studying gene function due to its relative ease of use and accuracy. Additionally, the system has enormous potential for treating hereditary diseases. Validation of CRISPR/Cas9 gene editing is necessary to ensure that genes of interest are successfully knocked down or knocked out. Here, we demonstrate how Molecular Devices' family of instruments can be utilized in gene editing experiments by using CRISPR/Cas9 to knockdown autophagy-related protein 5 (ATG5) in HEK293 cells. Cells successfully transfected with the CRISPR/Cas9 plasmid expressed GFP, and a 13% transfection efficiency was calculated automatically using the SpectraMax® i3x Multi-Mode Microplate Reader with MiniMax™ 300 Imaging Cytometer. The same platform was used to quantitate DNA and protein concentrations for subsequent validation experiments. We observed about a 70% decrease in ATG5 protein expression in CRISPR-edited cells compared to non-edited cells using the ScanLater™ Western Blot Detection System. Finally, autophagosome formation was measured in CRISPR-edited vs. non-edited cells via high-content imaging.
HOANG HA– Application Scientist, Molecular Devices, Sunnyvale, CA
I am an applications scientist at Molecular Devices, and I mainly support microplate readers. I have a M.S. in Biotechnology from UC Irvine and a B.S. in Cell Biology from UC Davis. My research background is based in Molecular and Cellular Biology. At UC Irvine, I studied stem cell regeneration in the mouse hair follicle, and before that I maintained stem cell cultures for hepatic differentiation at a stem-cell institute. I am interested in new biological technologies with potential therapeutic applications (ex. CRISPR). Additionally, I am still interested in all things molecular biology and cellular biology.