CRISPR-Cas9 All-in-one Plasmid

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

Design and clone gDNA under U6 promotor for gRNA transcription, the lentiviral compatible All-in-one plasmid expresses Cas-9 to facilitate the CRISPR targeting. EGFP is expressed from the same vector visualize transfection efficiency.

Materials Provided:	Vector: pLenti-EF1a-Cas9-EGFP-U6-sgRNA
Convenience Factors	- One vector to express Cas9 and gRNA - U6 promoter driving efficient gRNA transcription - EF1α promoter for high level Cas9 expression - Lentiviral compatible - EGFP for positive transfection identification
Vector Map
Brief Protocol	- Design your gene specific gDNA sequences - Clone gDNA sequences into pLenti-EF1a-Cas9-EGFP-U6-sgRNA - Transident transfection into cells for gene targeting - Visualize EGFP for transfection efficiency - Determine gene targeting success by your methods of choice