No enhancing effects of plasmid-specific histone acetyltransferase recruitment system on transgene expression in vivo.
No enhancing effects of plasmid-specific histone acetyltransferase recruitment system on transgene expression in vivo.
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质粒特异性组蛋白乙酰转移酶招募系统对体内转基因表达没有增强作用。
DOI：
10.1080/15257770.2019.1638514
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
M Fujimuro and Hiroyuki Kamiya
中科院分区：
文献类型：
--
作者：
T Suzuki;Y Wakao;T Watanabe;M Hori;Y Ikeda;H Tsuchiya;K Kogure;M Harada-Shiba;M Fujimuro and Hiroyuki Kamiya
组蛋白乙酰化水平的改变与染色体基因表达的变化有关。因此，结合在质粒DNA上的组蛋白的特异性乙酰化可能会增加转基因表达。以前，CREB结合蛋白的组蛋白乙酰转移酶结构域与GAL4序列依赖的DNA结合结构域(GAL4-HAT)的融合表达成功地提高了报告基因在培养细胞中的表达。Bioengng.123,277-280(2017)]。在这项研究中，同样的方法也应用于转基因小鼠的表达。将分别携带GAL4-HAT基因的激活子和报告基因的DNA经尾静脉注射入小鼠肝脏，检测小鼠血清中的赤霉素酶活性。出乎意料的是，共注射GAL4-HAT和荧光素酶质粒DNA似乎减少了荧光素酶的表达，而不是增加了。此外，联合注射明显减少了肝脏中荧光素酶DNA的数量。这些结果表明，该系统在体内是无效的，并建议排除表达GAL4-HAT的肝细胞。
Altered levels of histone acetylation are associated with changes in chromosomal gene expression. Thus, the specific acetylation of histones bound to plasmid DNA might increase transgene expression. Previously, the expression of the histone acetyltransferase domain of CREB-binding protein fused to the sequence-dependent DNA binding domain of GAL4 (GAL4-HAT) successfully improved reporter gene expression in cultured cells [J. Biosci. Bioengng.123, 277–280 (2017)]. In this study, the same approach was applied for transgene expression in mice. The activator and reporter plasmid DNAs bearing the genes for GAL4-HAT andGaussia princepsluciferase, respectively, were co-administered into the mouse liver by hydrodynamics-based tail vein injection, and theGaussialuciferase activity in serum was measured for two weeks. Unexpectedly, the co-injection of the GAL4-HAT and luciferase plasmid DNAs seemed to decrease, rather than increase, luciferase expression. Moreover, the co-injection apparently reduced the amount of luciferase DNA in the liver. These results indicated that this system is ineffective in vivo and suggested the exclusion of hepatic cells expressing GAL4-HAT.