Chen Ying-Jun, Chang Hao-Hsin, Lin Shih-Han, Lin Tzi-Yang, Wu Ting-Han, Lin Hsin-Ju, Liou Nan-Fu, Yang Chi-Jen, Chen Yuh-Tarng, Chang Kai Hsiang, Li Cen-You, Chou Ya-Hui
a Institute of Cellular and Organismic Biology, Academia Sinica , Taipei , Taiwan , ROC.
b Neuroscience Program of Academia Sinica , Taipei , Taiwan , ROC.
J Neurogenet. 2019 Mar-Jun;33(2):52-63. doi: 10.1080/01677063.2018.1564289. Epub 2019 Apr 2.
Several large or mid-scale collections of enhancer traps have been recently created to allow for genetic swapping of GAL4 coding sequences to versatile transcription activators or suppressors such as LexA, QF, split-GAL4 (GAL4-AD and GAL4-DBD), GAL80 and QS. Yet a systematic analysis of the feasibility and reproducibility of these tools is lacking. Here we focused on InSITE drivers that specifically label different subpopulations of olfactory neurons, particularly local interneurons (LNs), and genetically swapped the GAL4 domain for LexA, GAL80 or QF at the same locus. We found that the major utility-limiting factor for these genetic swaps is that many do not fully reproduce the original GAL4 expression patterns. Different donors exhibit distinct efficacies for reproducing original GAL4 expression patterns. The successfully swapped lines reported here will serve as valuable reagents and expand the genetic toolkits of olfactory circuit research.
最近已经创建了几个大型或中型的增强子陷阱文库，以实现GAL4编码序列与多种转录激活因子或抑制因子（如LexA、QF、分裂型GAL4（GAL4激活结构域和GAL4DNA结合结构域）、GAL80和QS）的基因交换。然而，目前缺乏对这些工具的可行性和可重复性的系统分析。在这里，我们聚焦于InSITE驱动子，它能特异性标记嗅觉神经元的不同亚群，特别是局部中间神经元（LN），并在同一基因座上对LexA、GAL80或QF进行GAL4结构域的基因交换。我们发现，这些基因交换的主要效用限制因素是许多交换不能完全重现原始的GAL4表达模式。不同的供体在重现原始GAL4表达模式方面表现出不同的效率。本文报道的成功交换的品系将作为有价值的试剂，并扩展嗅觉回路研究的基因工具包。
