Deep Intron Elements Mediate Nested Splicing Events at Consecutive AG Dinucleotides To Regulate Alternative 3′ Splice Site Choice in Vertebrate 4.1 Genes

Parra, Marilyn; Gallagher, Thomas L.; Amacher, Sharon L.; Mohandas, Narla; Conboy, John G.

doi:10.1128/mcb.05716-11

Cited by 17 publications

(28 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…CHIP-seq data indicate many potential splicing factor binding sites at long distances from intron/exon junctions (Yeo et al, 2009), but few specific examples of regulation-at-a-distance have been previously reported. These elements in the telomerase gene represent the first examples in mammals of specific distant sequences that influence splice choice by a mechanism other than introducing cryptic splice sites (Dominski and Kole, 1993; Friedman et al, 1999; Parra et al, 2012; Pros et al, 2009; Salem et al, 2012). …”

Section: Discussionmentioning

confidence: 99%

“…(E) Self-blast of human telomerase showing a block of repeats in intron 6 (block 6), a direct repeat in intron 6 (DR6), and a direct repeat in intron 8 (DR8). (F) Sequence logo created by WebLogo (Parra et al, 2012) showing the strong 38 nucleotide consensus sequence of the 26 repeats in block 6. (G) Diagram of the full minigene (c2) including block 6 (B6), the direct repeat in intron 6 (DR6), and the direct repeat in intron 8 (DR8).…”

Section: Highlightsmentioning

confidence: 99%

See 1 more Smart Citation

Regulation of Telomerase Alternative Splicing: A Target for Chemotherapy

Wong

Chen

Foster³

et al. 2013

Cell Reports

View full text Add to dashboard Cite

SUMMARY Telomerase is present in human cancer cells but absent in most somatic tissues. The mRNA of human telomerase (hTERT) is alternatively spliced into mostly non-functional products. We sought to understand splicing so we could decrease functional splice isoforms to reduce telomerase activity to complement direct enzyme inhibition. Unexpectedly, minigenes containing hTERT exons 5–10 flanked by 150–300bp intronic sequences did not produce alternative splicing. A 1.1kb region of 38bp repeats ~2kb from the exon 6/intron junction restored exclusion of exons 7/8. An element within intron 8, also >1kb from intron/exon junctions, modulated this effect. Transducing an oligonucleotide complementary to this second element increased non-functional hTERT mRNA from endogenous telomerase. These results demonstrate the potential of manipulating hTERT splicing for both chemotherapy and regenerative medicine, and provide the first specific sequences deep within introns that regulate alternative splicing in mammalian cells by mechanisms other than introducing cryptic splice sites.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Highlightsmentioning

confidence: 99%

Regulation of Telomerase Alternative Splicing: A Target for Chemotherapy

Wong

Chen

Foster³

et al. 2013

Cell Reports

View full text Add to dashboard Cite

show abstract

“…To date there are only twenty-eight studies using Vivo-morpholinos [17], [18], [19], [20], [21], [22], [23], [24], [25], [26], [27], [28], [29], [30], [31], [32], [33], [34], [35], [36], [37], [38], [39], [40], [41], [42], [43], [44] and all have reported at least 50% knockdown of the target gene with no adverse side effects. Fourteen of these studies used a mouse model [17], [18], [19], [20], [21], [24], [31], [32], [34], [35], [40], [41], [42], [43] with the remaining studies using rats [36], [37], newts [29], chicken embryos [27], fish [15], [16], [22], [23], [25], [26] tadpoles [30], [38], or frogs [28].…”

Section: Introductionmentioning

confidence: 99%

“…Fourteen of these studies used a mouse model [17], [18], [19], [20], [21], [24], [31], [32], [34], [35], [40], [41], [42], [43] with the remaining studies using rats [36], [37], newts [29], chicken embryos [27], fish [15], [16], [22], [23], [25], [26] tadpoles [30], [38], or frogs [28]. In the mouse models, it has been shown that Vivo-morpholinos were equally efficacious with IV or IP, and recent studies have shown success with direct injection in target tissue [36], [37].…”

Section: Introductionmentioning

confidence: 99%

Vivo-Morpholinos Induced Transient Knockdown of Physical Activity Related Proteins

2013

View full text Add to dashboard Cite

Physical activity is associated with disease prevention and overall wellbeing. Additionally there has been evidence that physical activity level is a result of genetic influence. However, there has not been a reliable method to silence candidate genes in vivo to determine causal mechanisms of physical activity regulation. Vivo-morpholinos are a potential method to transiently silence specific genes. Thus, the aim of this study was to validate the use of Vivo-morpholinos in a mouse model for voluntary physical activity with several sub-objectives. We observed that Vivo-morpholinos achieved between 60–97% knockdown of Drd1-, Vmat2-, and Glut4-protein in skeletal muscle, the delivery moiety of Vivo-morpholinos (scramble) did not influence physical activity and that a cocktail of multiple Vivo-morpholinos can be given in a single treatment to achieve protein knockdown of two different targeted proteins in skeletal muscle simultaneously. Knocking down Drd1, Vmat2, or Glut4 protein in skeletal muscle did not affect physical activity. Vivo-morpholinos injected intravenously alone did not significantly knockdown Vmat2-protein expression in the brain (p = 0.28). However, the use of a bradykinin analog to increase blood-brain-barrier permeability in conjunction with the Vivo-morpholinos significantly (p = 0.0001) decreased Vmat2-protein in the brain with a corresponding later over-expression of Vmat2 coincident with a significant (p = 0.0016) increase in physical activity. We conclude that Vivo-morpholinos can be a valuable tool in determining causal gene-phenotype relationships in whole animal models.

show abstract

“…41 Some twintron-like configurations can be resolved by a mechanism referred to as intrasplicing, but this has only been observed in the vertebrate 4.1R-and 4.1B paralogs, which encode cytoskeletal adaptor proteins. 15,42 This "twintron-like arrangement" may have evolved to facilitate an alternate splicing event that allows for a distant (downstream) exon to be incorporated into the mRNA. Specifically it involves the coordination of alternative splicing of an intron located in the 5 0 UTR region along with the utilization of an alternate promoter.…”

Section: Introductionmentioning

confidence: 99%