Efficient Generation of Genome-Modified Mice Using Campylobacter jejuni-Derived CRISPR/Cas

Area of Science:

• Genetics
• Molecular Biology
• Biotechnology

Background:

• Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas systems are vital for generating genome-modified animals.
• Existing CRISPR/Cas systems are widely used in mammalian zygote genome engineering.

Purpose of the Study:

• To investigate a novel CRISPR/Cas system derived from *Campylobacter jejuni* for mammalian zygote genome engineering.
• To assess the efficiency of this system in generating knockout and knock-in mouse models.

Main Methods:

• Utilized a *Campylobacter jejuni*-derived CRISPR/Cas system in mouse zygotes.
• Identified the protospacer-adjacent motif (5'-NNNVRYAC) recognized by this system.
• Performed zygote-mediated gene knockout and knock-in experiments.

Main Results:

• The *Campylobacter jejuni* CRISPR/Cas system efficiently generated knockout mice.
• This system successfully mediated zygote-mediated knock-in at a unique locus.
• The system demonstrated applicability in mammalian zygote genome engineering.

Conclusions:

• The novel bacterial CRISPR/Cas system is effective for generating knockout and knock-in mice.
• This system broadens the scope of zygote-mediated genome modification in animals.
• It offers a new tool for advancing the creation of genetically modified animal models.