Customized Mouse Models Created Using CRISPR-Cas9
GemPharmatech constructs more than 4000 customized mouse models per year utilizing CRISPR-Cas9 technology. This construction capacity ranks among the best in the world. CRISPR-Cas9 technology is the most common gene editing technology in use today and can be used to establish knockout mice (KO), conditional knockout mice (cKO), and knockin mice (KI).
Compared with other technologies, such as TALEN, ZFN, or ES, CRISPR-Cas9 is simple, fast, and low cost.
High quality: Create models with no random insertions and no serial recombination
High efficiency: Our platform results in a higher success rate and shorter delivery cycle than traditional methods
Multi-modification: Target multiple loci, multiple genes, and large fragments
Multi-background: Models can be created on a variety of genetic backgrounds, including C57BL/6J, C57BL/6N, BALB/c, FVB, NOD/LTJ, and NCG
Embryonic Stem Cell Homologous Recombination Technology for Creating Customized Mouse Models
Embryonic Stem Cell (ES) Homologous Recombination (HR) technology is a gene editing technique that integrates foreign DNA into the genome of ES cells. It is suitable for establishing mouse models with complex editing strategies or when the DNA inserts needed are larger than CRISPR-Cas9 technology can accommodate.
Self-deletion of resistance genes can save 3 months in model generation time
BAC shooting technology can achieve knock-in segments of 100-300kb
Well-established technique in use since 2005 with more than 700 models successfully completed
Models can be created on a variety of genetic backgrounds, including 129S6/SvEvTac and C57BL/6N
Creating Customized Mouse Models Using Transgenic Technology
Transgenic technology is generally used to establish customized models for overexpressed or conditionally overexpressed mice. In transgenic technology customization, DNA is randomly integrated into the mouse genome by injecting DNA directly into a fertilized mouse egg pronucleus using microinjection technology. With the increasing efficiency of knockins (KI), most models now use the H11 or Rosa26 site for KI to create overexpressed models.
|Comparison of transgenic technology and KI at the H11 or Rosa26 site|
F0 generation in as little as 8 weeks
F1 generation in as little as 16 weeks
Low technical difficulty
High technical difficulty
Different expression levels between founder lines, and expression levels over time (typically expression levels decrease)
Same expression levels in all founder lines, and constant expression levels over time
Random insertion site and number of copies
Accurate insertion location and gene copy number by design
Easy to alter or destroy the expression of other endogenous genes
Does not affect expression other endogenous genes
Poor genetic stability
High genetic stability
Large-scale work in establishing a stable line
Produces a stable line directly for analysis