

632617 · 1 mL
Takara · Cat: 632617
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takarabio.com/products/gene-function/gene-editing/crispr-cas...Description
The Gesicle Producer 293T Cell Line is a subclone of the transformed human embryonic kidney cell line, HEK 293, which is highly transfectable and supports high levels of protein expression. The cell line also constitutively expresses the simian virus 40 SV40 large T antigen. Gesicles are produced in these cells via co-overexpression of packaging mix components, which include a nanovesicle-inducing glycoprotein and a protein that is displayed on the cell surface and mediates binding and fusion with the cellular membrane of target cells. Simultaneous overexpression of another protein cargo can result in incorporation of that protein within the gesicles. When combined with a gesicle production system, these cells are capable of producing high gesicle yields. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back Cas9 protein delivery in culture cells.Immunohistochemistry was performed on RPE cells stably expressing ZsGreen1 and treated with Cas9 gesicles. Cells were stained 12 hr after addition of gesicles. Cas9 was detected using the Guide-it Cas9 Polyclonal Antibody Cat. # 632607 together with the Alexa 350-conjugated anti-rabbit IgG secondary antibody. Red fluorescence from the CherryPicker fluorescent protein could also be detected in the cells Back Successful knockout ofCD81in hiPS cells.Gesicles containing Cas9-sgRNA complexes designed to target human CD81 were harvested and added to Cellartis Human iPS Cell Line 18 hiPSC ChiPSC18 , cultured in Cellartis DEF-CS Culture System for 6 and 24 hr, and then cultured in gesicle-free DEF-CS culture media for an additional 7 days. The surface expression of CD81 on gesicle-treated cells and untreated control cells was determined via flow cytometry analysis using FITC-labeled antibodies against CD81. Panel A. CD81 negative left and positive right labeling controls with hiPSC ChiPSC18 cells. Panel B. DEF-hiPSC ChiPSC18 cells after 6 hr left and 24 hr right of gesicle treatment, labeled with anti-CD81 FITC antibodies.







