Generation of genetically modified rat models via the CRISPR/Cas9 technology

Area of Science:

• Molecular Biology
• Genetics
• Biotechnology

Background:

• The CRISPR/Cas9 system enables targeted genomic modification.
• This system comprises a single guide RNA (sgRNA) and Cas9 nuclease.
• Cas9 induces double-stranded DNA breaks (DSBs) at targeted genomic loci.

Purpose of the Study:

• To provide a detailed protocol for generating genetically modified rat models.
• To enable gene knockout or knockin in rats using CRISPR/Cas9 technology.

Main Methods:

• Micro-injection of the CRISPR/Cas9 system into rat embryos.
• Utilizing sgRNA to guide Cas9 nuclease to specific genomic sites.
• Leveraging DNA repair pathways (NHEJ or HDR) for gene editing.

Main Results:

• Successful generation of gene knockout rat models.
• Successful generation of gene knockin rat models.
• Demonstration of a robust protocol for CRISPR/Cas9 mediated genome editing in rats.

Conclusions:

• The described protocol facilitates efficient creation of gene knockout and knockin rat models.
• CRISPR/Cas9 technology offers a powerful tool for generating genetically modified rats for research.
• This method advances the development of novel disease models and genetic studies in rats.