CRISPR/Cas9 Knock-out Cell Line Services

Integrating exogenous plasmid DNA into the chromosome of host cells, and then express the target protein for a long time, which is called stable cell line. High-quality stable cell line plays an important role in biological research, including drug development, gene function research and recombinant antibody production. However, constructing an effective cell line is a complex and time-consuming task. KMD Bioscience has rich experience in stable cell line construction, our cell lines can be directly applied to downstream biological research. We have successfully constructed a large number of target cell lines for many research clients so far. Based on advanced technology and dedicated research teams, KMD Bioscience can provide the following services:

 

CRISPR/Cas9 Knock-out Cell Line Services

IOS Overexpression Stable Cell Line Services

IOS RNAi Stable Cell Line Services

Pinpoint Targeted Integration Cell Line Services

 

CRISPR/Cas9 Knock-out Cell Line Construction

CRISPR/Cas9 is a powerful technique for genome manipulation and causing a buzz in the science world. It has the potential to alter the course of research and drug discovery, by providing scientists with a powerful tool to change any gene, in any cell in a highly targeted manner and without introducing foreign DNA. In Type II CRISPR System (Fig 1), CRISPR RNA (crRNA) and trans-activating crRNA (tracrRNA) forms a complex through base pairing that can specifically recognize genomic sequences. Then the PAM (5'-NGG-3') sequence binds and invades DNA, directing the Cas9 endonuclease to generate double-strand breaks (DSBs) in the target fragment. This complex can be simplified by fusing crRNA and tracrRNA sequences to form single-guided RNA (sgRNA). The fused RNA has similar activity to wild-type and is more convenient for researchers because of its simplified structure. By connecting the elements of sgRNA and Cas9, it is possible to obtain a plasmid which can express both of them at the same time. Then it can be transfected into cells to manipulate the target gene.

 

 

Fig 1. Schematic representation of CRISPR/Cas9-mediated genome editing

 

 

 

 

Service Procedure:

Service Highlights:

*Customizable: We customize our offering to meet your specific project needs and provide experienced scientific support every step of the way.

*Extensive experience: Our experts are good at performing gene editing with CRISPR, from designing gRNA constructs for CRISPR to transfection and single clone generation of a wide range of cells.

*Higher knockout efficiency.

*We are able to knock out multiple genes on the same cell.

*We can provide a variety of expression vectors with different fluorescent and resistance markers, making it easier to screen for successful knock-out cells.