RRC ID 57141
著者 Raghu SV, Mohammad F, Chua JY, Lam JSW, Loberas M, Sahani S, Barros CS, Claridge-Chang A.
タイトル A zinc-finger fusion protein refines Gal4-defined neural circuits.
ジャーナル Mol Brain
Abstract 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.
巻・号 11(1)
ページ 46
公開日 2018-8-20
DOI 10.1186/s13041-018-0390-7
PII 10.1186/s13041-018-0390-7
PMID 30126464
PMC PMC6102859
MeSH Animals Drosophila Proteins / metabolism* Drosophila melanogaster / metabolism* Gene Expression Green Fluorescent Proteins / metabolism Nerve Net / metabolism* Protein Multimerization Recombinant Fusion Proteins / metabolism* Serotonergic Neurons / metabolism Transcription Factors / metabolism* Zinc Fingers*
IF 4.051
引用数 0
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