A Novel Single Vector Intersectional AAV Strategy for Interrogating Cellular Diversity and Brain Function.

Area of Science:

• Neuroscience
• Molecular Biology
• Genetics

Background:

• Advances in neuroscience require sophisticated tools to target specific cell populations in the brain based on complex criteria like gene expression and projection targets.
• Intersectional strategies enable conditional gene expression in cells where multiple features converge, but existing platforms have limitations.

Approach:

• Developed Conditional Viral Expression by Ribozyme Guided Degradation (ConVERGD), a single-construct intersectional targeting strategy.
• ConVERGD integrates a self-cleaving ribozyme with traditional FLEx switches for enhanced targeting capabilities.
• The ConVERGD vector design allows for easy modification, expanded intersectionality, and accommodation of larger payloads.

Key Points:

• ConVERGD offers a versatile platform for targeting diverse cell types with greater precision.
• Successfully employed ConVERGD to identify and target a novel subpopulation of norepinephrine (NE)-producing neurons in the rodent locus coeruleus (LC).
• These NE+ LC neurons co-express the prodynorphin (Pdyn) gene, a stress-related endogenous opioid.

Conclusions:

• ConVERGD is a powerful and adaptable tool for functional interrogation of neural circuitry.
• Targeted Pdyn-expressing NE+ LC neurons using ConVERGD.
• Demonstrated that this specific neuronal subpopulation can drive anxiogenic behavioral responses in rodents, highlighting its role in anxiety.