In the continuously evolving field of genetic research, the CRISPR/Cas9 system emerges as a groundbreaking technology for genetic modifications. This system affords researchers a multitude of genetic alterations, such as deletions, point mutations, and insertions. Remarkably, the CRISPR/Cas9 toolset provides substantial advantages over traditional methods, highlighting its efficiency, cost-effectiveness, and timeline advantages, especially when undertaking multiple gene modifications simultaneously.

What is CRISPR/Cas9?

The CRISPR/Cas9 technology employs a Cas9 nuclease equipped with two active sites. Upon binding to a guide RNA molecule, it generates targeted double-strand breaks in genomic DNA. Through these breaks, researchers can leverage donor DNA molecules, initiating homologous recombination to introduce specific nucleotide modifications into the target sequence.

CRISPR/Cas9 Platform

Creative Biogene’s CRISPR/Cas9 Platform specializes in generating transgenic mouse models that aid biomedical research and facilitate drug discovery. Offering a seamless service from experimental design to successful mouse model generation, the platform guarantees results in a competitive timeframe of three to four months. A commitment to delivering at least two founder mice or three F1 animals for knock-in projects underscores their dedication to quality and reliability.

Workflow

1. Strategy and Design: The process begins with a thorough assessment of the targeted gene sequence, involving target site selection, gRNA design, and donor DNA creation. Non-target site predictions also play a crucial role in ensuring precision.

2. CRISPR Vector Construction: This stage encompasses the synthesis and validation of gRNA and preparation of Cas9 mRNA, alongside constructing necessary DNA vectors.

3. Embryo Injection: The designed gRNA, Cas9 mRNA, and donor DNA are injected into mouse embryos to facilitate CRISPR-mediated modifications.

4. Founder Mice Production: Following embryo transplantation into pseudopregnant mice, screening via PCR and sequencing confirms successful insertions.

5. Breeding for F1 Mice: Depending on experimental requirements, founder mice may be bred with wild-type mice to generate progeny with the desired knock-in alleles.

For those targeting sequences under two hundred base pairs, donor DNA oligonucleotides are provided to facilitate the insertion of marker stop regions or functional elements. Conversely, for larger sequences exceeding two hundred base pairs, donor vectors are utilized, fostering the integration of luminescent or fluorescent reporter genes at precise genomic locations.

Key Benefits

• High efficiency with gRNA, achieving near-total Cas9 target cutting efficiency.
• Proven strategies for large segment knock-ins.
• Assurance of germline transmission through careful founder breeding.
• Timely updates on project progress.
• Efficient transportation services for samples.

Diverse Applications

The CRISPR/Cas9 knock-in mouse models offer invaluable insights across various research domains, including:

• Examination of gene and protein biological functions.
• Quantitative assessments of gene expression levels.
• Exploration of transcriptional regulatory elements.
• Construction of models for specific human diseases.
• Drug development and screening methodologies.
• Preclinical safety assessments and toxicity analyses.
• Validation of drug targets and genomic verification of antibodies.

Creative Biogene remains committed to providing outstanding CRISPR/Cas9 gene editing services. They prioritize detailed communication with clients to keep them informed on project evolution. Alongside state-of-the-art laboratories, animal care compliant with stringent standards, and a team of experienced scientists, Creative Biogene aims to fulfill unique research demands, safeguarding the value of scientific endeavors.