While CRISPR/Cas9-mediated gene editing is a powerful technique for genome manipulation, two significant challenges remain: obtaining efficient delivery of Cas9 and the gene-specific single guide RNA (sgRNA) to all cell types, and achieving fewer off‑target effects. To address these challenges, we have developed cell‑derived nanovesicles called gesicles that are created using the Guide-it CRISPR/Cas9 Gesicle Production System. CRISPR/Cas9 gesicles contain active Cas9 protein complexed with an sgRNA specific to a gene of interest, and mediate binding and fusion with the membrane of a wide range of target cells.
While CRISPR/Cas9-mediated gene editing is a powerful technique for genome manipulation, two significant challenges remain: obtaining efficient delivery of Cas9 and the gene-specific single guide RNA (sgRNA) to all cell types, and achieving fewer off‑target effects. To address these challenges, we have developed cell‑derived nanovesicles called gesicles that are created using the Guide-it CRISPR/Cas9 Gesicle Production System. CRISPR/Cas9 gesicles contain active Cas9 protein complexed with an sgRNA specific to a gene of interest, and mediate binding and fusion with the membrane of a wide range of target cells. These features can increase gene-editing efficiency to levels that are often higher than those obtained using plasmid‑based delivery methods. Additionally, this method allows for tight control of the dose and duration of the Cas9‑sgRNA complex in the cell, with the added benefit of reduced off‑target effects. The nanovesicles are produced in a Gesicle Producer 293T Cell Line via co-overexpression of packaging-mix components, including a nanovesicle-inducing glycoprotein and a protein for cell-surface display that mediates binding and fusion with the cellular membrane of target cells. Simultaneous overexpression of 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, they will efficiently enter the cells and mediate efficient gene editing.
This system provides the components needed for cloning and expressing your target-specific guide RNA, plus packaging reagents for producing CRISPR/Cas9 gesicles. The Gesicle Producer 293T Cell Line is sold separately.
美国Clontech公司是一家专门致力于基因工程及蛋白质工程领域研究用产品的研发和生产的专业公司。公司产品涉及PCR产品、RT-PCR产品、cDNA文库构建、基因克隆、基因表达、基因分析、蛋白质技术等诸多领域。产品种类多达2000多种。Clontech公司拥有多种独立知识产权产品,其强大的科研实力和完善的质量管理体系(ISO9001)为Clontech产品的良好品质提供了可靠保障。