Silencing gene expression by targeting chromosomal DNA with antigene peptide nucleic acids and duplex RNAs

被引:31
作者
Janowski, Bethany A. [1 ]
Hu, Jiaxin [1 ]
Corey, David R. [1 ]
机构
[1] Univ Texas Dallas, SW Med Ctr, Dept Biochem Pharmacol, Dallas, TX 75390 USA
关键词
D O I
10.1038/nprot.2006.64
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The value of recognizing cellular RNA sequences by short interfering RNAs (siRNAs) in mammalian cells is widely appreciated, but what might be learned if it were also possible to recognize chromosomal DNA? Recognition of chromosomal DNA would have many applications, such as inhibiting gene expression, activating gene expression, introducing mutations, and probing chromosome structure and function. We have shown that antigene peptide nucleic acids (agPNAs) and antigene duplex RNAs (agRNAs) block gene expression and probe chromosomal DNA. Here we describe a protocol for designing antigene agents and introducing them into cells. This protocol can also be used to silence expression with PNAs or siRNAs that target mRNA. From preparation of oligomers to analysis of data, experiments with agPNAs and agRNAs require similar to 14 d and 9 d, respectively.
引用
收藏
页码:436 / 443
页数:8
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