CRISPR-Gene Specific Duo Set for Knock In (KI)
CRISPR-Cas (Clustered regularly-interspaced short palindromic repeats) has evolved to be the prokaryotic immune system that confers resistance to foreign genetic elements such as plasmids and phages, and provides a form of acquired immunity. The CRISPR-Cas9 system has been adapted in mammalian system for targeted gene knock-out (KO) and Knock-in (KI).
The CRISPR system is "cheap, quick and easy to use" (Nature, 2015, Vol 522, pages 20-24) for editing gene features in mammalian cells, Lifeome configured CRISPR-Cas9 systems to make it even easier to adopt in your lab by offering:
- All-in-one Plasmid with Cas9 readily expressed, just clone your gDNA
- Ready to use, gene specific pre-designed Duo-gRNA sets, just transfect target cells
- Pre-designed gene specific Duo-gRNA sets for in vitro gRNA production
- AAV and Lentiviral CRISPR systems available
- Mouse and Rat knock-out and knock-in services, constitutive or inducible
For most human, mouse and rat genes, Lifeome has selected two gRNA targeting sequences and cloned into pGL3-U6 gRNA transcription vector. The sequences are selected for high effiency targeting using CRISPR-Cas9 system but low "off-target" potentials. The Duo Sets can be co-transfected with Cas9 expression plasmids into cells for effective gene targeting.
Target gene knock-in (KI) is achieved by through homologous recombination. A donor plasmid containing both left and right arms of a homologous sequences to the Cas9 nicking site, flanking a GFP-Puromycin cassette that can be replaced with a gene of interest. After the plasmids for Cas9 expression, gRNA expression and the donor plasmid are delivered into the cells, the following three steps will happen sequentially to set the GFP-Puromycin cassette into the target genomic location:
1. Cas9-gRNA complex nicks the intended location;
2. Homologous recombination mechanism will "patch" the nick with the GFP-Puromycin cassette
MCP 1 Rat$3,078.00
