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小鼠活跃长散在核元件1亚家族定量PCR检测方法的建立及其在衰老相关逆转座分析中的应用

Establishment of Quantitative PCR Assays for Active Long Interspersed Nuclear Element-1 Subfamilies in Mice and Applications to the Analysis of Aging-Associated Retrotransposition.

作者信息

Kuroki Ryota, Murata Yui, Fuke Satoshi, Nakachi Yutaka, Nakashima Jun, Kujoth Gregory C, Prolla Tomas A, Bundo Miki, Kato Tadafumi, Iwamoto Kazuya

机构信息

Department of Molecular Brain Science, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan.

Lab for Molecular Dynamics of Mental Disorders, RIKEN Center for Brain Science, Wako, Japan.

出版信息

Front Genet. 2020 Sep 16;11:519206. doi: 10.3389/fgene.2020.519206. eCollection 2020.

Abstract

The retrotransposon long interspersed nuclear element-1 (LINE-1) can autonomously increase its copy number within a host genome through the retrotransposition process. LINE-1 is active in the germline and in neural progenitor cells, and its somatic retrotransposition activity has a broad impact on neural development and susceptibility to neuropsychiatric disorders. The method to quantify the genomic copy number of LINE-1 would be important in unraveling the role of retrotransposition, especially in the brain. However, because of the species-specific evolution of LINE-1 sequences, methods for quantifying the copy number should be independently developed. Here, we developed a quantitative PCR (qPCR) assay to measure the copy number of active LINE-1 subfamilies in mice. Using the assay, we investigated aging-associated alterations of LINE-1 copy number in several brain regions in wild-type mice and mice as a model for accelerated aging. We found that aged mice showed higher levels of the type GfII LINE-1 in the basal ganglia than the wild-type mice did, highlighting the importance of assays that focus on an individual active LINE-1 subfamily.

摘要

逆转录转座子长散在核元件1(LINE-1)可通过逆转录转座过程在宿主基因组内自主增加其拷贝数。LINE-1在生殖系和神经祖细胞中具有活性,其体细胞逆转录转座活性对神经发育和神经精神疾病易感性具有广泛影响。在阐明逆转录转座的作用时,尤其是在大脑中,定量LINE-1基因组拷贝数的方法将具有重要意义。然而,由于LINE-1序列的物种特异性进化,应独立开发定量拷贝数的方法。在此,我们开发了一种定量PCR(qPCR)检测方法来测量小鼠中活跃LINE-1亚家族的拷贝数。使用该检测方法,我们研究了野生型小鼠和作为加速衰老模型的小鼠几个脑区中与衰老相关的LINE-1拷贝数变化。我们发现,老年小鼠基底神经节中GfII型LINE-1的水平高于野生型小鼠,这突出了专注于单个活跃LINE-1亚家族的检测方法的重要性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/df7e/7525186/70e2e4b9e97b/fgene-11-519206-g001.jpg

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