Inhibiting gene expression with locked nucleic acids (LNAs) that target chromosomal DNA
Inhibiting gene expression with locked nucleic acids (LNAs) that target chromosomal DNA
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DOI：
10.1021/bi700227g
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发表时间：
2007-06-26
期刊：
影响因子：
2.9
通讯作者：
Corey, David R.
中科院分区：
文献类型：
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作者：
Beane, Randall L.;Ram, Rosalyn;Corey, David R.
含有锁核酸碱基（LNA）的寡核苷酸对互补DNA序列具有增加的亲和力。我们假设增强的亲和力可能允许LNA识别人类细胞内的染色体DNA并抑制基因表达。为了验证这一假设，我们合成了与孕酮受体（PR）和雄激素受体（AR）启动子内序列互补的抗原LNA（agLNA）。我们观察到AR和PR表达的抑制agLNAs，但没有类似的寡聚体含有2 '-甲氧基乙基碱或noncomplementary LNA。抑制是剂量依赖性的，并且表现出< 10 nM的IC 50值。有效的抑制依赖于agLNA的长度、LNA碱基的位置、LNA取代的数量和靶序列在靶向启动子内的位置。靶向转录起始位点处或附近序列的LNA比靶向转录因子结合位点或反向重复序列的LNA产生更好的抑制。这些结果表明agLNA可以识别染色体靶序列并有效地阻断基因表达。agLNA可用于基因沉默、作为染色体结构的细胞探针和治疗应用。
Oligonucleotides containing locked nucleic acid bases (LNAs) have increased affinity for complementary DNA sequences. We hypothesized that enhanced affinity might allow LNAs to recognize chromosomal DNA inside human cells and inhibit gene expression. To test this hypothesis, we synthesized antigene LNAs (agLNAs) complementary to sequences within the promoters of progesterone receptor (PR) and androgen receptor (AR). We observed inhibition of AR and PR expression by agLNAs but not by analogous oligomers containing 2'-methoxyethyl bases or noncomplementary LNAs. Inhibition was dose dependent and exhibited IC50 values of < 10 nM. Efficient inhibition depended on the length of the agLNA, the location of LNA bases, the number of LNA substitutions, and the location of the target sequence within the targeted promoter. LNAs targeting sequences at or near transcription start sites yielded better inhibition than LNAs targeting transcription factor binding sites or an inverted repeat. These results demonstrate that agLNAs can recognize chromosomal target sequences and efficiently block gene expression. agLNAs could be used for gene silencing, as cellular probes for chromosome structure, and therapeutic applications.