Genomic engineering in cell lines is a versatile tool for studying gene function, designing diseases models, biopharmaceutical research, drug discovery and many other applications. CRISPR (Clustered Regulatory Interspaced Short Palindromic Repeats)/Cas9 systems is a newly developed yet the most popular method for genome editing. It has been widely used in current biology, functional genome screening, cell-based human hereditary disease modeling, epigenomic studies and visualization of cellular processes.
CRISPR/Cas9 system consists of a “guide” RNA (gRNA) and a bacterial CRISPR-associated endonuclease (Cas9). The gRNA is a short synthetic RNA composed of a Cas9-binding “scaffold” sequence and ∼20 nucleotide “targeting” sequence that defines the target genomic site to be modified. Cas9 contains two nuclease domain to induce site-specific DNA cleavage. Besides the basic employment of gene knock-out, CRISPR/Cas9 system can be applied to selective gene activation and repression, purification of specific genomic DNA and fluorescent protein-tagging in live cells through modifications of the Cas9. It’s also a scalable genome-wide editing technology for its ease of generating gRNAs. The simplicity and high-efficiency of CRISPR/Cas9 system make it a preferable genomic engineering method to the traditional ZNF and TALEN system.
Figure Basic gene editing with CRISPR/Cas9 System
Creative Biogene is providing new services using CRISPR/Cas9 System to make various forms of engineered cell lines. We have years of experience in generating the following cell lines.
• Knockout cell lines
• Conditional Knockout cell lines
• Site-specific knock-in cell lines
• Gene Mutation cell lines
• Double gene knockout cell lines
• Gene tagging cell lines
Our services include:
• Construct generation
Targeting vector construction and confirmation, and select the most efficient engineered nuclease
• Stable cell clones selection
Perform transfection on target cells including HEK293 cells, CHO cells, CHO D44 cells, HeLa cells, other tumor cells, Mouse ES cells, Human ES cells, Primary cells etc. Select cell by MACS, FACS and screen gene-edited cell clones. Expand and cryopreserve the engineered cells.
• Gene editing evaluation
Evaluate the target stable cell clones via one or several assays such as Western Blot, ELISA, real-time PCR, or reporter assays etc.