Cell Proliferation and Cell Cycle Analysis using a Multiple Laser Microplate Cytometer
Poster May 03, 2007
Andrew Goulter, Paul Wylie and Wayne Bowen
Fluorescence microplate cytometers are heavily used in oncology research including cell proliferation and cell cycle analysis using the DNA stain propidium iodide (488 nm excitation). Here, we describe the extension of this capability through the use of DNA stains, Hoechst 34580 (405 nm) and TO-PRO®-3 (633 nm) on an Acumen eX3, and discuss the potential for multiplexing in immunodetection assays.
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE