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Related Experiment Videos

PNA interference mapping demonstrates functional domains in the noncoding RNA Xist.

A Beletskii1, Y K Hong, J Pehrson

  • 1Harvard Institute of Human Genetics, Beth Israel Deaconess Medical Center, Harvard Medical School, 4 Blackfan Circle, Boston, MA 02115, USA.

Proceedings of the National Academy of Sciences of the United States of America
|August 2, 2001
PubMed
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Xist RNA binding is essential for X-chromosome inactivation. Inhibiting Xist

Area of Science:

  • Epigenetics
  • Molecular Biology
  • Genetics

Background:

  • The noncoding RNA Xist is crucial for X-chromosome inactivation.
  • Xist coats the inactive X-chromosome (Xi), but its binding necessity for inactivation is unclear.

Purpose of the Study:

  • To determine if Xist binding to the X-chromosome is required for X-chromosome inactivation.
  • To functionally map the chromatin-binding region of Xist RNA.

Main Methods:

  • Developed and utilized peptide nucleic acid (PNA) interference mapping to study noncoding RNA function in living cells.
  • Employed cell-permeating PNA targeted against specific regions of Xist RNA.

Main Results:

  • Inhibition of Xist association with the X-chromosome led to the failure of X-chromosome silencing.

Related Experiment Videos

  • A single 19-bp antisense PNA targeting Xist RNA disrupted the inactive X-chromosome (Xi).
  • PNA interference mapping also disturbed the association of Xi with macro-histone H2A.

    • Conclusions:

    • Xist binding to the X-chromosome is necessary for initiating and maintaining X-chromosome inactivation.
    • Peptide nucleic acid interference mapping is an effective method for studying noncoding RNA function in vivo.