{"product_id":"flat-tagged-crispr-cas9-nuclease-rgd-fiber-modified-adenovirus-adrgd-gfp-flag-hcas9-bhv21600434","title":"FLAT tagged CRISPR\/Cas9 nuclease RGD-fiber modified Adenovirus (Ad(RGD)-GFP-FLAG-hCas9)","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\u003cp\u003eAd(RGD)-GFP-FLAG-hCas9 is a replication-defective recombinant Ad5 adenovirus expressing the GFP-FLAG-hCas9 nuclease under the CMV promoter. It is intended for genome editing in difficult-to-transfect cell types and in vivo, typically paired with a separate gRNA-delivery vector or co-delivered gRNA cassette.\u003c\/p\u003e\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\u003cul\u003e\n\u003cli\u003e\n\u003cstrong\u003eBackbone:\u003c\/strong\u003e Human adenovirus type 5 (Ad5) with E1 and E3 deleted (dE1\/E3). Replication-incompetent in standard cells; replication-competent helper cells (HEK293) are required for amplification.\u003c\/li\u003e\n\u003cli\u003e\n\u003cstrong\u003ePromoter (CMV):\u003c\/strong\u003e a strong, ubiquitous promoter active in most mammalian cell types.\u003c\/li\u003e\n\u003cli\u003e\n\u003cstrong\u003eTransgene:\u003c\/strong\u003e GFP-FLAG-hCas9 (eGFP tag).\u003c\/li\u003e\n\u003cli\u003e\n\u003cstrong\u003eTiter \u0026amp; format:\u003c\/strong\u003e 1×10\u003csup\u003e10\u003c\/sup\u003e PFU\/ml in storage buffer (DMEM, 2% BSA, 2.5% glycerol or equivalent), supplied as a 200 µL aliquot.\u003c\/li\u003e\n\u003c\/ul\u003e\u003ch2\u003eBiological background\u003c\/h2\u003e\u003cp\u003eThe Type II prokaryotic CRISPR\/Cas system is the new class of tools for targeted genome engineering. The cas9 nucleases derived from clustered regularly interspaced short palindromic repeats (CRISPR)-cas systems uses small RNAs as guides (gRNA) to cleave DNA in a sequence-specific manner. With its ease in designing guide sequences to target specific genomic loci, the CRISPR\/Cas system is a much simpler, faster, and robust alternative to TALEN and Zinc finger nuclease platforms.\u003c\/p\u003e\u003cp\u003eFor site-specific genome editing, the CRISPR\/Cas9 system requires the Cas 9 nuclease and the gRNA. This RGD-fiber modified adenovirus expresses a codon-optimized spCas9 nuclease with FLAG tag and a GFP reporter. It can be used along with guided RNA for genome-editing in cells that regular Ad5 doesn’t infect well, including human or mouse macrophages, ES cells, T cells, synoviocytes, certain fibroblast and islet grafts etc.\u003c\/p\u003e\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\u003cul\u003e\n\u003cli\u003eDecision-relevant for researchers studying CRISPR\/Cas9 Editing.\u003c\/li\u003e\n\u003cli\u003eAdenovirus-mediated delivery is well-established in primary cells, organoids, and small-animal models.\u003c\/li\u003e\n\u003c\/ul\u003e\u003ch2\u003eCommon research applications\u003c\/h2\u003e\u003cul\u003e\n\u003cli\u003eTargeted gene knockout in difficult-to-transfect cells, paired with gRNA delivery.\u003c\/li\u003e\n\u003cli\u003eIn vivo somatic genome editing via tissue-targeted adenoviral delivery.\u003c\/li\u003e\n\u003c\/ul\u003e\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\u003cul\u003e\n\u003cli\u003eAdenoviral delivery is episomal and non-integrating; expression dilutes with cell division and typically lasts 1–2 weeks in dividing cells (longer in non-dividing cells such as hepatocytes, neurons, and cardiomyocytes).\u003c\/li\u003e\n\u003cli\u003ePre-existing anti-Ad5 neutralizing antibodies are common in human and primate hosts and can reduce in vivo transduction; this is less relevant in inbred laboratory mouse strains.\u003c\/li\u003e\n\u003cli\u003eMOI optimization is essential — over-dosing can cause cytopathic effects; under-dosing yields incomplete transduction. A 3–5× MOI titration in your specific cell or animal model is recommended.\u003c\/li\u003e\n\u003cli\u003eReplication-defective Ad5 vectors are typically handled at BSL-2; consult your institutional biosafety officer for specific transgenes and routes of use.\u003c\/li\u003e\n\u003c\/ul\u003e\u003c!-- Sources (internal):\n  - NCBI Gene: https:\/\/www.ncbi.nlm.nih.gov\/gene\n  - UniProt: https:\/\/www.uniprot.org\/\n  - Russell WC. Adenoviruses: update on structure and function. J Gen Virol 2009; 90:1–20.\n  - Alba R, Bosch A, Chillon M. Gutless adenovirus: last-generation adenovirus for gene therapy. Gene Ther 2005; 12 Suppl 1:S18–27.\n  - Jinek M et al. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 2012; 337:816–21.\n  - Cong L et al. Multiplex genome engineering using CRISPR\/Cas systems. Science 2013; 339:819–23.\n  - Vendor reference: https:\/\/www.vectorbiolabs.com\/product\/1905-flat-tagged-crispr-cas9-nuclease-rgd-fiber-modified-adenovirus\/\n--\u003e","brand":"Vector Biolabs","offers":[{"title":"1x10^10 PFU\/ml \/ 200 µL","offer_id":53286502826349,"sku":"1905","price":690.0,"currency_code":"USD","in_stock":true}],"url":"https:\/\/www.ebiohippo.com\/products\/flat-tagged-crispr-cas9-nuclease-rgd-fiber-modified-adenovirus-adrgd-gfp-flag-hcas9-bhv21600434","provider":"BioHippo","version":"1.0","type":"link"}