Inhibition of mycobacterium smegmatis gene expression and growth using antisense peptide nucleic acids
Inhibition of mycobacterium smegmatis gene expression and growth using antisense peptide nucleic acids
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DOI：
10.1159/000088840
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发表时间：
2005-01-01
影响因子：
1.2
通讯作者：
Good, L
中科院分区：
文献类型：
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作者：
Kulyté, A;Nekhotiaeva, N;Good, L
在mRNA水平上抑制基因的反义药物是全基因组研究和药物靶标验证的有吸引力的工具。这种方法可能特别适合研究难以用标准遗传工具操作的细菌。带有载体肽的反义肽核酸（PNA）可以抑制大肠杆菌和金黄色葡萄球菌的基因表达。在这里，我们询问肽- pnas是否可以介导耻垢分枝杆菌的反义作用。我们首先针对绿色荧光蛋白报告基因，并观察到低微摩尔浓度下的剂量和序列依赖性抑制。PNA和靶mRNA序列的序列改变消除了抑制作用，有力地支持了一种反义抑制机制。此外，反义PNAs与各种附加肽表现出更好的抗gfp作用。两个靶向ha必需基因的肽pnas抑制生长，并引起类似于ha缺失细胞的细胞形态变化。因此，反义肽- pnas可以有效和特异性地抑制分枝杆菌的报告基因和内源性必需基因。版权所有(C) 2005 S. Karger AG，巴塞尔。
Antisense agents that inhibit genes at the mRNA level are attractive tools for genome-wide studies and drug target validation. The approach may be particularly well suited to studies of bacteria that are difficult to manipulate with standard genetic tools. Antisense peptide nucleic acids (PNA) with attached carrier peptides can inhibit gene expression in Escherichia coli and Staphylococcus aureus. Here we asked whether peptide-PNAs could mediate antisense effects in Mycobacterium smegmatis. We first targeted the gfp reporter gene and observed dose- and sequence-dependent inhibition at low micromolar concentrations. Sequence alterations within both the PNA and target mRNA sequences eliminated inhibition, strongly supporting an antisense mechanism of inhibition. Also, antisense PNAs with various attached peptides showed improved anti-gfp effects. Two peptide-PNAs targeted to the essential gene inhA were growth inhibitory and caused cell morphology changes that resemble that of InhA-depleted cells. Therefore, antisense peptide-PNAs can efficiently and specifically inhibit both reporter and endogenous essential genes in mycobacteria. Copyright (C) 2005 S. Karger AG, Basel.