An Animal Model for Genetic Analysis of Multi-Gene Families: Cloning and Transgenesis of Large Tandemly Repeated Histone Gene Clusters.
An Animal Model for Genetic Analysis of Multi-Gene Families: Cloning and Transgenesis of Large Tandemly Repeated Histone Gene Clusters.
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多基因家族遗传分析的动物模型：大型串联重复组蛋白基因簇的克隆和转基因。
DOI：
10.1007/978-1-4939-8663-7_17
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Matera,AGregory
中科院分区：
文献类型：
--
作者：
Meers,MichaelP;Leatham-Jensen,Mary;Penke,TaylorJR;McKay,DanielJ;Duronio,RobertJ;Matera,AGregory
组蛋白翻译后修饰（PTM）被认为参与了一系列重要的分子和细胞过程，包括基因表达，复制和核组织。重要的是，组蛋白PTM也被认为是跨细胞周期和世代的表观遗传信息载体的主要候选者。然而，由于动物基因组中组蛋白基因的高度重复性，通过诱变直接测试组蛋白PTM本身在这些过程中的必要性是极其困难的。我们开发了一个转基因系统，以产生果蝇基因型，其中复制依赖性组蛋白基因的整个互补序列在感兴趣的残基处突变。我们构建了BAC载体，其含有用于谱系追踪的可见标记，沿着具有克隆大（60-100 kb）插入物的能力，所述插入物随后可以位点特异性地整合到D中。黑胃基因组我们证明，人工串联阵列的核心5 kb的复制依赖组蛋白重复可以产生相对容易。该遗传平台代表了第一个利用单个串联转基因插入进行分子和细胞表型遗传学和分析的组蛋白替代系统。我们证明了我们的系统的效用，直接防止组蛋白残基被修改，并研究随之而来的表型。该系统可以推广到任何生物学目的串联重复序列的克隆和转基因插入。
Histone post-translational modifications (PTMs) are thought to participate in a range of essential molecular and cellular processes, including gene expression, replication, and nuclear organization. Importantly, histone PTMs are also thought to be prime candidates for carriers of epigenetic information across cell cycles and generations. However, directly testing the necessity of histone PTMs themselves in these processes by mutagenesis has been extremely difficult to carry out because of the highly repetitive nature of histone genes in animal genomes. We developed a transgenic system to generateDrosophila melanogastergenotypes in which the entire complement of replication-dependent histone genes is mutant at a residue of interest. We built a BAC vector containing a visible marker for lineage tracking along with the capacity to clone large (60–100 kb) inserts that subsequently can be site-specifically integrated into theD. melanogastergenome. We demonstrate that artificial tandem arrays of the core 5 kb replication-dependent histone repeat can be generated with relative ease. This genetic platform represents the first histone replacement system to leverage a single tandem transgenic insertion for facile genetics and analysis of molecular and cellular phenotypes. We demonstrate the utility of our system for directly preventing histone residues from being modified, and studying the consequent phenotypes. This system can be generalized to the cloning and transgenic insertion of any tandemly repeated sequence of biological interest.
影响因子：
11.8
作者：
McKay, Daniel J.;Klusza, Stephen;Penke, Taylor J. R.;Meers, Michael P.;Curry, Kaitlin P.;McDaniel, Stephen L.;Malek, Pamela Y.;Cooper, Stephen W.;Tatomer, Deirdre C.;Lieb, Jason D.;Strahl, Brian D.;Duronio, Robert J.;Matera, A. Gregory
通讯作者：
Matera, A. Gregory
影响因子：
5.4
作者：
Carlson SM;Gozani O
通讯作者：
Gozani O
影响因子：
30.8
作者：
Park, JH;Cosgrove, MS;Boeke, JD
通讯作者：
Boeke, JD