Neural circuit-specific gene manipulation in mouse brain in vivo using split-intein-mediated split-Cre system

Area of Science:

• Neuroscience
• Molecular Biology
• Genetics

Background:

• Analyzing neural circuit function in vivo requires efficient genetic manipulation tools.
• Current methods may lack the specificity or versatility needed for complex circuit studies.

Purpose of the Study:

• To develop and describe an advanced circuit-selective gene manipulating tool for in vivo neural circuit analysis.
• To enable visualization and manipulation of specific neural circuits using a novel viral strategy.

Main Methods:

• Development of anterograde and retrograde adeno-associated viruses (AAVs).
• Encoding split-intein-mediated split-Cre for circuit-selective gene manipulation.
• Production and purification of AAVs, viral injection into the mouse brain, and imaging analysis.

Main Results:

• A novel strategy for circuit-selective gene manipulation in vivo was successfully developed.
• The tool allows for both visualization and manipulation of genes within specific neural circuits.
• The protocol details AAV production, delivery, and subsequent imaging analysis.

Conclusions:

• The developed AAV-based system provides an efficient and advanced method for studying neural circuits.
• This tool facilitates the analysis of individual neural circuit functions and gene manipulation in vivo.
• The described protocol offers a valuable resource for neuroscience research requiring precise circuit targeting.