A spliced intron accumulates as a lariat in the nucleus of T cells

Qian, Lian; Vu, Minh N.; Carter, Mark; Wilkinson, Miles

doi:10.1093/nar/20.20.5345

Cited by 75 publications

(65 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Another factor that limits debranching is the intrinsic stability of the branched lariat. Certain introns can accumulate as lariats within the nucleus despite functional DBR1 (Qian et al 1992). A study of Pem homeobox gene introns in HeLa cells suggests that intron half-lives are z9-29 min, the shortest being only 9.4 min, and thus intron degradation is rate-limited by the lariat debranching step rather than exonuclease activity (Clement et al 2001).…”

Section: Dmpk-mirt 5 Knockdown Is Dependent On Splicing and Exportin-mentioning

confidence: 99%

Artificial mirtron-mediated gene knockdown: Functional DMPK silencing in mammalian cells

Seow¹,

Sibley²,

Wood³

2012

RNA

View full text Add to dashboard Cite

Mirtrons are introns that form pre-miRNA hairpins after splicing to produce RNA interference (RNAi) effectors distinct from Drosha-dependent intronic miRNAs. Here we present a design algorithm for artificial mirtrons and demonstrate, for the first time, efficient gene knockdown of myotonic dystrophy protein kinase (DMPK) target sequences in Renilla luciferase 39 UTR and subsequently pathogenic DMPK mRNA, causative of Type I myotonic dystrophy, using artificial mirtrons cloned as eGFP introns. Deep sequencing of artificial mirtrons suggests that functional mature transcripts corresponding to the designed sequence were produced in high abundance. They were further shown to be splicing-dependent, Drosha-independent, and partially dependent on exportin-5, resulting in the precise generation of pre-miRNAs. In a murine myoblast line containing a pathogenic copy of human DMPK with more than 500 CUG repeats, the DMPK artificial mirtron corrected DM1-associated splicing abnormalities of the Serca-1 mRNA, demonstrating the therapeutic potential of mirtron-mediated RNAi. Thus, further development and exploitation of the unique properties of mirtrons will benefit future research and therapeutic RNAi applications as an alternative to conventional RNAi strategies.

show abstract

Section: Dmpk-mirt 5 Knockdown Is Dependent On Splicing and Exportin-mentioning

confidence: 99%

Artificial mirtron-mediated gene knockdown: Functional DMPK silencing in mammalian cells

Seow¹,

Sibley²,

Wood³

2012

RNA

View full text Add to dashboard Cite

show abstract

“…2; Zhang et al 2013). Such stable lariat introns were first described on a gene by gene basis in the early 1990s and include the nuclear accumulation of introns from the Delta transcript in Drosophila (Kopczynski and Muskavitch 1992) and the T cell receptor-β mRNA in T cells (Qian et al 1992). More recently, deep sequencing has revealed that a subset of introns is strikingly over-represented and presumably has features blocking the normal degradation pathway of debranching and exonuclease digestion.…”

Section: Circular Rna From Intronsmentioning

confidence: 99%

Circular RNAs: diversity of form and function

Lasda

Parker

2014

RNA

1,016

818

View full text Add to dashboard Cite

It is now clear that there is a diversity of circular RNAs in biological systems. Circular RNAs can be produced by the direct ligation of 5 ′ and 3 ′ ends of linear RNAs, as intermediates in RNA processing reactions, or by "backsplicing," wherein a downstream 5 ′ splice site (splice donor) is joined to an upstream 3 ′ splice site (splice acceptor). Circular RNAs have unique properties including the potential for rolling circle amplification of RNA, the ability to rearrange the order of genomic information, protection from exonucleases, and constraints on RNA folding. Circular RNAs can function as templates for viroid and viral replication, as intermediates in RNA processing reactions, as regulators of transcription in cis, as snoRNAs, and as miRNA sponges. Herein, we review the breadth of circular RNAs, their biogenesis and metabolism, and their known and anticipated functions.

show abstract

“…This would influence the results only if the resulting lariat is very stable. Generally, spliced intron sequences are believed to be rapidly degraded (Sharp, 1994), but isolated examples exist where lariats accumulate in the nucleus (Quian et al, 1992). Considering the precipitous decline in template molecules after day 5, however, it is apparent that the unknown half-life of the primary transcript and the spliced lariat can be neglected in the data analysis, as both must be much shorter than that of the mRNA.…”

Section: Discussionmentioning

confidence: 99%

Transcriptional regulation of FABP expression in flight muscle of the desert locust, Schistocerca gregaria

Zhang

Haunerland

1998

Insect Biochemistry and Molecular Biology

View full text Add to dashboard Cite

FABP is the most abundant cytosolic protein in developed flight muscle of adult locusts, but it is completely absent in nymphs. During the two weeks following adult ecdysis, FABP rises to 18% of the total soluble proteins. Its mRNA increases steadily up to day 8, before it gradually declines until reaching a low concentration at day 15, which remains constant for the rest of the animal's life. Using a PCR primer combination specific for a 597 bp sequence of intron 1, we have developed a reverse transcription PCR assay to quantify the amount of primary transcript present in muscle tissue at various ages. The FABP gene is not transcribed prior to metamorphosis; its primary transcript rises rapidly during the first two days of adult life, and remains close to maximal until day 5. Subsequently its level rapidly declines, ultimately reaching values of less than 0.02% of the maximal level. The correlation between primary transcript, mRNA and FABP content were analyzed by modeling transcription, translation and degradation with computer modeling software. The computer simulation is in good agreement with the experimentally obtained data, suggesting that the control of FABP expression in locust flight muscle occurs predominantly at the level of transcription initiation.

show abstract

A spliced intron accumulates as a lariat in the nucleus of T cells

Cited by 75 publications

References 23 publications

Artificial mirtron-mediated gene knockdown: Functional DMPK silencing in mammalian cells

Artificial mirtron-mediated gene knockdown: Functional DMPK silencing in mammalian cells

Circular RNAs: diversity of form and function

Transcriptional regulation of FABP expression in flight muscle of the desert locust, Schistocerca gregaria

Contact Info

Product

Resources

About