Manipulating the Prion Protein Gene Sequence and Expression Levels with CRISPR/Cas9

Area of Science:

• Neuroscience
• Genetics
• Molecular Biology

Background:

• The mammalian prion protein (PrP) is central to prion diseases and implicated in Alzheimer's disease, cancer, and stroke.
• Studying PrP function requires genetic manipulation, but the Prnp locus is challenging to modify using traditional methods like homologous recombination.

Purpose of the Study:

• To explore the utility of CRISPR/Cas9 (CC9) genome engineering for manipulating the endogenous mouse Prnp locus.
• To demonstrate the feasibility of modifying Prnp sequences and enhancing PrP expression using CC9-based synergistic activation mediators (SAMs).

Main Methods:

• Systematic application of the CRISPR/Cas9 (CC9) system to target the endogenous mouse Prnp locus.
• Utilizing CC9-based synergistic activation mediators (SAMs) to modulate PrP expression.

Main Results:

• Successful modification of the mouse Prnp locus using CRISPR/Cas9 (CC9) technology.
• Demonstrated ability to boost PrP expression via CC9-mediated SAMs.

Conclusions:

• CRISPR/Cas9 (CC9) offers powerful new possibilities for manipulating the Prnp locus, overcoming previous technical hurdles.
• These gene-targeting techniques can aid researchers in studying prion diseases and normal PrP functions in transgenic mouse models and cell culture.