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

Autor: Raghu SV; Program in Neuroscience and Behavioral Disorders, Duke-NUS Medical School, Singapore, 138673, Singapore.; Agency for Science Technology and Research, Institute for Molecular and Cell Biology, Singapore, Singapore.; Present address: Neurogenetics lab, Mangalore University, Mangalore, Karnataka, India., Mohammad F; Program in Neuroscience and Behavioral Disorders, Duke-NUS Medical School, Singapore, 138673, Singapore.; Agency for Science Technology and Research, Institute for Molecular and Cell Biology, Singapore, Singapore., Chua JY; Program in Neuroscience and Behavioral Disorders, Duke-NUS Medical School, Singapore, 138673, Singapore.; Agency for Science Technology and Research, Institute for Molecular and Cell Biology, Singapore, Singapore., Lam JSW; Agency for Science Technology and Research, Institute for Molecular and Cell Biology, Singapore, Singapore., Loberas M; Agency for Science Technology and Research, Institute for Molecular and Cell Biology, Singapore, Singapore., Sahani S; Agency for Science Technology and Research, Institute for Molecular and Cell Biology, Singapore, Singapore., Barros CS; Plymouth University Peninsula Schools of Medicine and Dentistry, Plymouth, PL6 8BU, UK., Claridge-Chang A; Program in Neuroscience and Behavioral Disorders, Duke-NUS Medical School, Singapore, 138673, Singapore. adamcc@gmail.com.; Agency for Science Technology and Research, Institute for Molecular and Cell Biology, Singapore, Singapore. adamcc@gmail.com.; Department of Physiology, National University of Singapore, Singapore, Singapore. adamcc@gmail.com.
Jazyk: angličtina
Zdroj: Molecular brain [Mol Brain] 2018 Aug 20; Vol. 11 (1), pp. 46. Date of Electronic Publication: 2018 Aug 20.
DOI: 10.1186/s13041-018-0390-7
Abstrakt: 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.
Databáze: MEDLINE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje