Functional Domains of NEAT1 Architectural lncRNA Induce Paraspeckle Assembly through Phase Separation

Mol Cell. 2018 Jun 21;70(6):1038-1053.e7. doi: 10.1016/j.molcel.2018.05.019.


A class of long noncoding RNAs (lncRNAs) has architectural functions in nuclear body construction; however, specific RNA domains dictating their architectural functions remain uninvestigated. Here, we identified the domains of the architectural NEAT1 lncRNA that construct paraspeckles. Systematic deletion of NEAT1 portions using CRISPR/Cas9 in haploid cells revealed modular domains of NEAT1 important for RNA stability, isoform switching, and paraspeckle assembly. The middle domain, containing functionally redundant subdomains, was responsible for paraspeckle assembly. Artificial tethering of the NONO protein to a NEAT1_2 mutant lacking the functional subdomains rescued paraspeckle assembly, and this required the NOPS dimerization domain of NONO. Paraspeckles exhibit phase-separated properties including susceptibility to 1,6-hexanediol treatment. RNA fragments of the NEAT1_2 subdomains preferentially bound NONO/SFPQ, leading to phase-separated aggregates in vitro. Thus, we demonstrate that the enrichment of NONO dimers on the redundant NEAT1_2 subdomains initiates construction of phase-separated paraspeckles, providing mechanistic insights into lncRNA-based nuclear body formation.

Keywords: 1,6-hexanediol; CRISPR/Cas9; RNA-binding proteins; architectural RNA; haploid cell; noncoding RNA; nuclear bodies; paraspeckle; phase separation; prion-like domain.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • CRISPR-Cas Systems
  • Cell Nucleus / metabolism
  • HeLa Cells
  • Humans
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Protein Domains
  • RNA Recognition Motif Proteins / genetics
  • RNA Recognition Motif Proteins / metabolism
  • RNA Stability
  • RNA, Long Noncoding / genetics*
  • RNA, Long Noncoding / metabolism*
  • Transcription Factors / metabolism


  • NEAT1 long non-coding RNA, human
  • Nuclear Proteins
  • RNA Recognition Motif Proteins
  • RNA, Long Noncoding
  • Transcription Factors