CRISPR/Cas9 Knockin Mice: Transforming Genetic Research

The CRISPR/Cas9 technology has revolutionized genetic modification, enabling precise alterations in the genome of various organisms. Among its numerous applications, the creation of knockin mice represents a significant advancement in biomedical research, providing invaluable tools for studying gene function and modeling human diseases.

Understanding the CRISPR/Cas9 System

The CRISPR/Cas9 system is engineered to induce targeted double-strand breaks (DSBs) in DNA, using a guide RNA (gRNA) that directs the Cas9 nuclease to specific genomic locations. This capability permits the introduction of precise modifications, including point mutations and larger insertions that are critical for the study of specific genes. By employing donor DNA during the repair of DSBs, researchers can insert desired sequences into target genes, thus creating knockin mouse models tailored for specific research needs.

Comprehensive Services at Creative Biogene

Creative Biogene specializes in offering extensive services to facilitate the generation of CRISPR/Cas9 knockin mice. From the initial strategy development to the successful production of founder mice, every step is meticulously managed by our expert team. Our commitment is to provide high efficiency and fast turnaround, generally delivering at least two founders or three F1 animals within a three to four-month timeframe.

The Workflow

1. Strategy and Design: Tailored strategies are developed based on the target gene sequence. This includes nuclease-mediated strategy services such as gRNA design, donor DNA design, and non-target prediction.

2. CRISPR Vector Construction: gRNA is synthesized and validated while preparing Cas9 mRNA. We construct the necessary DNA vectors to facilitate the gene modification.

3. Injection into Mouse Embryos: Both gRNA and Cas9 mRNA, alongside donor DNA, are injected into mouse embryos ensuring an adequate number for successful outcomes.

4. Production of Founder Mice: The injected embryos are implanted into pseudopregnant mice, with the offspring verified by PCR and sequencing for successful genetic alterations.

5. Breeding for F1 Generation: Depending on experimental goals, the founder mice may be bred to yield F1 mice, allowing for further analysis of gene function in a consistent genetic background.

In cases where a target sequence is limited to less than 200 base pairs, we provide oligonucleotides as donor DNA, suitable for inserting key regulatory elements or marker sequences. For larger sequences, donor vectors are utilized, facilitating the addition of reporter genes which can enable in-depth study of gene expression.

Benefits of Our Service

• High efficiency of gRNA ensuring nearly complete target cutting.
• Well-established protocols that support substantial knockin operations.
• Assured germline transmission through F1 breeding.
• Regular updates on project progress and efficient communication.
• Reliable and convenient transport services for genetic materials.

Applications of Knockin Mice

Knockin mice serve diverse applications in research and development:

• Exploration of gene and protein functions.
• Quantitative studies on gene expression.
• Examination of transcriptional regulatory elements.
• Development of specific models for human diseases.
• Drug discovery and evaluation of therapeutic options.
• Verification of drug targets and their specificity.

A Professional Commitment to Quality

Creative Biogene is dedicated to delivering professional CRISPR/Cas9 gene editing services with an emphasis on quality and reliability. Our advanced laboratories, expert team, and animal care standards ensure that your research objectives are met with precision. Our goal is to support every step of your project from conception to completion, protecting your scientific investments.