Functional dissection of NEAT1 using genome editing reveals substantial localization of the NEAT1-1 isoform outside paraspeckles
- Publication Type:
- Journal Article
- RNA, 2017, 23 (6), pp. 872 - 881
- Issue Date:
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© 2017 Li et al. Large numbers of long noncoding RNAs have been discovered in recent years, but only a few have been characterized. NEAT1 (nuclear paraspeckle assembly transcript 1) is a mammalian long noncoding RNA that is important for the reproductive physiology of mice, cancer development, and the formation of subnuclear bodies termed paraspeckles. The two major isoforms of NEAT1 (3.7 kb NEAT1-1 and 23 kb NEAT1-2 in human) are generated from a common promoter and are produced through the use of alternative transcription termination sites. This gene structure has made the functional relationship between the two isoforms difficult to dissect. Here we used CRISPR-Cas9 genome editing to create several different cell lines: total NEAT1 knockout cells, cells that only express the short form NEAT1-1, and cells with twofold more NEAT1-2. Using these reagents, we obtained evidence that NEAT1-1 is not a major component of paraspeckles. In addition, our data suggest NEAT1-1 localizes in numerous nonparaspeckle foci we termed "microspeckles," which may carry paraspeckle-independent functions. This study highlights the complexity of lncRNA and showcases how genome editing tools are useful in dissecting the structural and functional roles of overlapping transcripts.
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