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果蝇 attP40 背景对 Or47b 嗅觉受体神经元肾小球组织的影响。

The effect of Drosophila attP40 background on the glomerular organization of Or47b olfactory receptor neurons.

机构信息

Department of Biology, Duke University, Durham, NC 27708, USA.

出版信息

G3 (Bethesda). 2023 Apr 11;13(4). doi: 10.1093/g3journal/jkad022.

Abstract

Bacteriophage integrase-directed insertion of transgenic constructs into specific genomic loci has been widely used by Drosophila community. The attP40 landing site located on the second chromosome gained popularity because of its high inducible transgene expression levels. Here, unexpectedly, we found that homozygous attP40 chromosome disrupts normal glomerular organization of Or47b olfactory receptor neuron (ORN) class in Drosophila. This effect is not likely to be caused by the loss of function of Msp300, where the attP40 docking site is inserted. Moreover, the attP40 background seems to genetically interact with the second chromosome Or47b-GAL4 driver, which results in a similar glomerular defect. Whether the ORN phenotype is caused by the neighbouring genes around Msp300 locus in the presence of attP40-based insertions or a second unknown mutation in the attP40 background remains elusive. Our findings tell a cautionary tale about using this popular transgenic landing site, highlighting the importance of rigorous controls to rule out the attP40 landing site-associated background effects.

摘要

噬菌体整合酶将转基因构建物插入特定基因组位点的方法已被果蝇研究界广泛应用。位于第二号染色体上的 attP40 着陆点因其可诱导的转基因表达水平高而受到青睐。然而,令人意外的是,我们发现纯合 attP40 染色体破坏了果蝇中 Or47b 嗅觉受体神经元 (ORN) 类的正常肾小球组织。这种影响不太可能是由于 attP40 对接位点插入的 Msp300 功能丧失引起的。此外,attP40 背景似乎与第二号染色体 Or47b-GAL4 驱动子在遗传上相互作用,导致类似的肾小球缺陷。在存在基于 attP40 的插入的情况下,ORN 表型是否是由 Msp300 基因座周围的邻近基因引起的,或者是 attP40 背景中的第二个未知突变引起的,目前仍不清楚。我们的研究结果告诫人们要谨慎使用这种流行的转基因着陆点,强调需要严格控制以排除 attP40 着陆点相关的背景效应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/38cf/10085800/0f74edce8cbe/jkad022f1.jpg

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