$1,155.00Catalog Number: CRISPR-Duo-KISize: kits

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

Gene Specific gRNA Duo Sets for Gene Knock In

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

Materials Provided: Two clones of pGL3-U6 with gene specific gDNA cloned
One donor clone with genomic sequences flanking GFP-Puromycin cassette for homologous recombination.
Materials Needed: A Cas-9 expression clone is needed (such as Purchase pST1374-NLS-Cas9 OR Purchase pST1374-NLS-Cas9-EGFP
How to Order 1. Search the gene you want to knock out, by UniProt ID, or gene name;
2. Add CRISPR-Duo to cart, and note the UniProt ID on check out.
3. Our product specialist will contact you for specifics of gene of interest and cell line to be used.
Vector Map CRISPR-gRNA expression vector

 

Quantity

$1,155.00

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