title: Phosphorothioate oligonucleotides can displace NEAT1 RNA and form nuclear paraspeckle-like structures.
abstract: Nuclear paraspeckles are built co-transcriptionally around a long non-coding RNA, NEAT1. Here we report that transfected 20-mer phosphorothioate-modified (PS) antisense oligonucleotides (ASOs) can recruit paraspeckle proteins to form morphologically normal and apparently functional paraspeckle-like structures containing no NEAT1 RNA. PS-ASOs can associate with paraspeckle proteins, including P54nrb, PSF, PSPC1 and hnRNPK. NEAT1 RNA can be displaced by transfected PS-ASO from paraspeckles and rapidly degraded. Co-localization of PS-ASOs with P54nrb was observed in canonical NEAT1-containing paraspeckles, in perinucleolar caps upon transcriptional inhibition, and importantly, in paraspeckle-like or filament structures lacking NEAT1 RNA. The induced formation of paraspeckle-like and filament structures occurred in mouse embryonic stem cells expressing little or no NEAT1 RNA, suggesting that PS-ASOs can serve as seeding molecules to assemble paraspeckle-like foci in the absence of NEAT1 RNA. Moreover, CTN, an RNA reported to be functionally retained in paraspeckles, was also observed to localize to paraspeckle-like structures, implying that paraspeckle-like structures assembled on PS-ASOs are functional. Together, our results indicate that functional paraspeckles can form with short nucleic acids other than NEAT1 RNA.
authors.affiliation: Department of Core Antisense Research, ISIS Pharmaceuticals, Inc. 2855 Gazelle Court, Carlsbad, CA 92010, USA.
authors.forename: Wen
authors.initials: W
authors.lastname: Shen
authors.affiliation: Department of Core Antisense Research, ISIS Pharmaceuticals, Inc. 2855 Gazelle Court, Carlsbad, CA 92010, USA Lliang@isisph.com.
authors.forename: Xue-hai
authors.initials: XH
authors.lastname: Liang
authors.affiliation: Department of Core Antisense Research, ISIS Pharmaceuticals, Inc. 2855 Gazelle Court, Carlsbad, CA 92010, USA.
authors.forename: Stanley T
authors.initials: ST
authors.lastname: Crooke
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D022003:Cell Nucleus Structures / Q000737:chemistry*; D002478:Cells, Cultured; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D009687:Nuclear Proteins / Q000032:analysis; D016376:Oligonucleotides, Antisense / Q000032:analysis* / Q000378:metabolism; D054735:Phosphorothioate Oligonucleotides / Q000032:analysis* / Q000378:metabolism; D062085:RNA, Long Noncoding / Q000378:metabolism*; D014162:Transfection
chemical_list: C577276:NEAT1 long non-coding RNA, human; C577275:NEAT1 long non-coding RNA, mouse; D009687:Nuclear Proteins; D016376:Oligonucleotides, Antisense; D054735:Phosphorothioate Oligonucleotides; D062085:RNA, Long Noncoding