Guide-it™ CRISPR/Cas9 Gesicle Production System

The Guide-it CRISPR/Cas9 Gesicle Production System is a system for producing high yields of target-specific CRISPR/Cas9 gesicles for gene editing. Gesicles are cell-derived nanovesicles used to deliver macromolecular cargoes to a broad range of target cells, including cells that are difficult to transfect with plasmids. The nanovesicles are produced in a Gesicle Producer 293T Cell Line (Cat. No. 632617) via co-overexpression of packaging mix components, which include a nanovesicle-inducing glycoprotein and a protein that is displayed on the cell surface that mediates binding and fusion with the cellular membrane of target cells. Simultaneous overexpression of additional macromolecular cargoes, in this case the Cas9 protein from Streptococcus pyogenes and a target-specific guide RNA (sgRNA), results in incorporation of the Cas9/sgRNA complex within the gesicles. After the resulting Cas9/sgRNA gesicles are harvested and applied to your target cells in the presence of protamine sulfate, they will efficiently enter the cells and mediate gene editing. This system provides the components needed to clone and express your target-specific guide RNA, and packaging reagents to produce CRISPR/Cas9 gesicles. It is essential to use the pGuide-it-sgRNA1 Vector for expression and packaging of your guide RNA, since using other commonly used guide RNA vectors will not result in effective Cas9/sgRNA complexes.