CD BioGlyco is a professional company providing Glycobiology Disease Model Development Services to clients such as Glycobiology Disease Model Construction, Custom In Vivo Glycobiology Disease Model, In Vivo Glycobiology Disease Model Screening, and In Vitro Glycobiology Disease Model Screening services. Our lab uses a specialized clustered regularly interspaced short palindromic repeat sequence (CRISPR) system to generate glycobiological disease models. Genome editing based on the CRISPR-CRISPR associated protein 9 (CRISPR/Cas9) system precisely introduces mutations in the target DNA sequence. This accelerates and advances the creation of customized animal models.
Based on advanced CRISPR systems, we construct genome-editing animal models using microinjections, electroporation, and genome editing via oviductal nucleic acid delivery (GONAD). CD BioGlyco offers a wide range of gene editing services including point mutations (substitutions), small sequence insertions, knockin, large deletion, and chromosomal rearrangements (translocations, duplications, and inversions). We provide not only the common plasmid-based CRISPR/Cas9 but also CRISPR/Cas13, Cas9 mRNA/gRNA, and Cas9/gRNA ribonucleoproteins (RNPs). Based on the characteristics of the client's model, our researchers recommend the most suitable construction method for you. Our researchers achieve high-efficiency targeting by inducing double-strand breaks at the expected locus via CRISPR/Cas. Meanwhile, we provide in vivo enhancer characterization services.
CD BioGlyco mainly provides the following several services:
CD BioGlyco provides the CRISPR/Cas9 system as a powerful gene editor for efficient and high-quality production of gene knockout mice and rats. Our researchers co-inject Cas9 mRNA and enhanced green fluorescent protein (EGFP) sgRNA into transgenic animals to achieve CRISPR/Cas9-mediated EGFP knockout. Common knockout genes include, but are not limited to ATP6V1H gene, Notch3 gene, Fah gene, Hpd gene, Asl genes, SOX9 gene, Irx3 gene, Irx5 genes, p53 gene, LKB1 gene, KRAS gene and so on. We offer double, triple, or multiple knockout services depending on client needs.
One or more nucleotides in the genome of a point mutant mouse and rat are replaced by a variant nucleotide. This may result in in-frame amino acid changes or code-shifting mutations in the protein sequence. With the support of CRISPR/Cas9, we use DNA double-strand break (DSB)-induced homologous recombination (HR)-dependent or independent repair mechanisms to generate point mutation mouse and rat. We obtain conditional mutant animals by co-injection of zygotes with Cas9 mRNA, different guide RNAs (sgRNAs), and DNA vectors of different sizes.
Based on homology-directed repair (HDR), our researchers induce precise large-fragment knockin service at the single nucleotide level. We inject HDR-targeted embryonic stem (ES) cells into wild-type host blastocysts to generate knockin mouse and rat strains. Our researcher achieves a one-step generation of knockin mouse and rat for targeted genes by injecting Cas9 mRNA in combination with sgRNA and single-stranded DNA oligonucleotide complexes/constructs into the cytoplasm of a zygote.
With the high-quality CRISPR/Cas9 system, we generate knockdown mice models by inducing insertion deletions that cause shifted codons. CRISPR-induced insertion deletions at the junction of exons and introns caused selective splicing and generated a wide variety of different mRNAs. Our researchers introduce these mRNAs into premature termination codons, leading to decreased expression of specific genes.
Technology: Plasmid construction, Transcription, Microinjection, PCR amplification, Agarose gel electrophoresis, T7EI assay, PCR sequencing, Cloning and Sanger sequencing, Western blot
Journal: Scientific Reports
Published: 2016
IF: 4.6
Results: In this work, researchers used truncated RNA-guided nucleases (tru-RGNs) to microinject zygote-generating Factor VII (FVII) gene-knockout mice with a high efficiency of up to 80.1%. By comparing the two approaches of standard RGNs (std-RGNs) and tru-RGNs, the researchers found that the ratio of the first site to the second site was significantly higher in tru-gRNAs than in std-gRNAs. Notably, the off-target frequency of knockout mice was much lower than that of the cell line through both tru-RGNs and std-RGNs-mediated gene editing.
Fig.2 Analysis of RGN-induced genomic mutations in NIH/3T3 cells. (An, et al., 2016)
CD BioGlyco attaches great importance to the cycle time and quality of our services and is committed to providing the most satisfactory solutions for our clients. Our operators all have rich backgrounds in gene editing. We are committed to promoting the development of our client projects. Please feel free to contact us.
References
