The infectious BAC genomic DNA expression library: a high capacity vector system for functional genomics.
The infectious BAC genomic DNA expression library: a high capacity vector system for functional genomics.
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DOI：
10.1038/srep28644
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发表时间：
2016-06-29
影响因子：
4.6
通讯作者：
Wade-Martins R
中科院分区：
文献类型：
--
作者：
Lufino MM;Edser PA;Quail MA;Rice S;Adams DJ;Wade-Martins R
基因剂量在一系列细胞表型中起关键作用，然而大多数细胞表达系统使用基于异源cDNA的载体，其表达的蛋白质远高于生理水平。相比之下，基因组DNA表达载体通过携带包括其调控元件的基因的整个基因组DNA基因座来产生生理相关水平的基因表达。在这里，我们描述了第一个基因组DNA表达文库产生的高容量单纯疱疹病毒-1扩增子技术提供细菌人工染色体（BAC）到细胞中的病毒转导。感染性BAC（iBAC）文库含有184，320个克隆，平均插入片段大小为134.5kb。我们在中国仓鼠卵巢（CHO）疾病模型细胞系和小鼠胚胎干（ES）细胞中显示，该文库可用于在一系列背景下的遗传拯救研究，包括Ldlr缺陷的生理恢复和病毒受体表达。iBAC库代表了一个重要的新的遗传分析工具，可供研究界公开使用。
Gene dosage plays a critical role in a range of cellular phenotypes, yet most cellular expression systems use heterologous cDNA-based vectors which express proteins well above physiological levels. In contrast, genomic DNA expression vectors generate physiologically-relevant levels of gene expression by carrying the whole genomic DNA locus of a gene including its regulatory elements. Here we describe the first genomic DNA expression library generated using the high-capacity herpes simplex virus-1 amplicon technology to deliver bacterial artificial chromosomes (BACs) into cells by viral transduction. The infectious BAC (iBAC) library contains 184,320 clones with an average insert size of 134.5 kb. We show in a Chinese hamster ovary (CHO) disease model cell line and mouse embryonic stem (ES) cells that this library can be used for genetic rescue studies in a range of contexts including the physiological restoration of Ldlr deficiency, and viral receptor expression. The iBAC library represents an important new genetic analysis tool openly available to the research community.
在插入插入的BLM缺陷型胚胎干细胞库中逆转录病毒感染所需的宿主因子的隐性遗传筛查。
影响因子：
12.3
作者：
Wang, Wei;Bradley, Allan
通讯作者：
Bradley, Allan