PNA interference mapping demonstrates functional domains in the noncoding RNA Xist.
The noncoding RNA Xist has been shown to be essential for X-chromosome inactivation and to coat the inactive X-chromosome (Xi). Thus, an important question in understanding the formation of Xi is whether the binding reaction of Xist is necessary for X-chromosome inactivation. In this article, we demonstrate the failure of X-chromosome silencing if the association of Xist with the X-chromosome is inhibited. The chromatin-binding region was functionally mapped and evaluated by using an approach for studying noncoding RNA function in living cells that we call peptide nucleic acid (PNA) interference mapping. In the reported experiments, a single 19-bp antisense cell-permeating PNA targeted against a particular region of Xist RNA caused the disruption of the Xi. The association of the Xi with macro-histone H2A is also disturbed by PNA interference mapping.
Beletskii
A
A
Harvard Institute of Human Genetics, Beth Israel Deaconess Medical Center, Harvard Medical School, 4 Blackfan Circle, Boston, MA 02115, USA.
Hong
Y K
YK
Pehrson
J
J
Egholm
M
M
Strauss
W M
WM
D000818:Animals; D001483:Base Sequence; D005260:Female; D006657:Histones / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D020135:Peptide Nucleic Acids / Q000737:chemistry*; D012313:RNA / Q000737:chemistry / Q000378:metabolism*; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated / Q000737:chemistry* / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism; D014960:X Chromosome
D006657:Histones; D020135:Peptide Nucleic Acids; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated; D014157:Transcription Factors; C077885:XIST non-coding RNA; D012313:RNA