
The authors have developed highly efficient conditional gene manipulation systems, including GFP-RNAi/Flp-out and CRISPR/Cas9-based approaches, to target all chemical transmission (chemoconnectome, CCT) genes in Drosophila. These tools enable detailed dissection of neural circuits underlying specific behaviors.


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Dissection of neural circuitry underlying behaviors is a central theme in neurobiology. We have previously proposed the concept of chemoconnectome (CCT) to cover the entire chemical transmission between neurons and target cells in an organize and created tools for studying it (CCTomics) by targeting all genes related to the CCT in Drosophila. Here we have created lines targeting the CCT in conditional manners after modifying GFP RNA interference, Flp-out and CRISPR/Cas9 technologies. All three strategies are validated to be highly effective with the best using chromatin-peptide fused Cas9 variants and scaffold optimized sgRNAs. As a proof of principle, we conduct a comprehensive intersection analysis of CCT genes expression profiles in the clock neurons, uncovering 43 CCT genes present in clock neurons. Specific elimination of each from clock neurons revealed that loss of the neuropeptide CNMa in two posterior dorsal clock neurons (DN1ps) or its receptor (CNMaR) caused advanced morning activity, indicating a suppressive role of CNMa-CNMaR on morning anticipation, opposite to the promoting role of PDF-PDFR on morning anticipation. These results demonstrate the effectiveness of conditional CCTomics and its tools created by us here and establish an antagonistic relationship between CNMa-CNMaR and PDF-PDFR signaling in regulating morning anticipation.

### Competing Interest Statement

The authors have declared no competing interest.

Conditional Chemoconnectomics (cCCTomics): Conditional Targeting of Chemical Transmission Efficiently

conditional-chemoconnectomics-a-highly-efficient-toolkit-for-targeted-manipulation-of-chemical-transmission-genes-in-drosophila


Conditional Chemoconnectomics: A Highly Efficient Toolkit for Targeted Manipulation of Chemical Transmission Genes in Drosophila
