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A zinc-finger fusion protein refines Gal4-defined neural circuits

By Shamprasad Varija Raghu, Farhan Mohammad, Jia Yi Chua, Claudia Barros, Joanne Lam, Mavis Loberas, Sadhna Sahani, Adam Claridge-Chang

Posted 04 Dec 2017
bioRxiv DOI: 10.1101/228718 (published DOI: 10.1186/s13041-018-0390-7)

The analysis of behavior requires that the underlying neuronal circuits are identified and genetically isolated. In several major model species -- most notably Drosophila, neurogeneticists identify and isolate neural circuits with a binary heterologous expression-control system: Gal4-UASG. One limitation of Gal4-UASG is that expression patterns are often too broad to map circuits precisely. To help refine the range of Gal4 lines, we developed an intersectional genetic AND operator. Interoperable with Gal4, the new system's key component is a fusion protein in which the DNA-binding domain of Gal4 has been replaced with a zinc finger domain with a different DNA-binding specificity. In combination with its cognate binding site (UASZ) the zinc-finger-replaced Gal4 ('Zal1') was functional as a standalone transcription factor. Zal1 transgenes also refined Gal4 expression ranges when combined with UASGZ, a hybrid upstream activation sequence. In this way, combining Gal4 and Zal1 drivers captured restricted cell sets compared with single drivers and improved genetic fidelity. This intersectional genetic AND operation presumably derives from the action of a heterodimeric transcription factor: Gal4-Zal1. Configurations of Zal1-UASZ and Zal1-Gal4-UASGZ are versatile tools for defining, refining, and manipulating targeted neural expression patterns with precision.

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