Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (172K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Nasim, Md. T.
Right arrow Articles by Eperon, I. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Nasim, Md. T.
Right arrow Articles by Eperon, I. C.
Related Collections
Right arrow RNA characterisation and manipulation
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2002, Vol. 30, No. 20 e109
© 2002 Oxford University Press

A double reporter assay for detecting changes in the ratio of spliced and unspliced mRNA in mammalian cells

Md. Talat Nasim, H. M. Chowdhury and I. C. Eperon*

Department of Biochemistry, University of Leicester, University Road, Leicester LE1 7RH, UK

*To whom correspondence should be addressed. Tel: +44 116 2523482; Fax: +44 116 2523369; Email: eci{at}leicester.ac.uk

Current methods for measuring the efficiency of splicing in mammalian cells rely on either direct analysis of the RNA, which does not lend itself to rapid assays, or on single reporter functions that are subject to numerous intrinsic variables. If two protein activities are encoded within a single reading frame but on separate exons, with an intervening sequence containing termination codons, then the expression of the second activity is dependent on removal of the intervening sequence by pre-mRNA splicing. Thus, the ratio of the activities encoded by exon 2 to exon 1 reflects the ratio of expression from spliced mRNA to the total expression of spliced and unspliced RNA. This provides a rapid and convenient assay for the effects on splicing efficiency of trans-acting factors or of alterations in the sequences of the intron and surrounding exon sequences.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Hum Mol GenetHome page
M. T. Nasim, A. Ghouri, B. Patel, V. James, N. Rudarakanchana, N. W. Morrell, and R. C. Trembath
Stoichiometric imbalance in the receptor complex contributes to dysfunctional BMPR-II mediated signalling in pulmonary arterial hypertension
Hum. Mol. Genet., June 1, 2008; 17(11): 1683 - 1694.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
P. Loyer, J. H. Trembley, J. A. Grenet, A. Busson, A. Corlu, W. Zhao, M. Kocak, V. J. Kidd, and J. M. Lahti
Characterization of Cyclin L1 and L2 Interactions with CDK11 and Splicing Factors: INFLUENCE OF CYCLIN L ISOFORMS ON SPLICE SITE SELECTION
J. Biol. Chem., March 21, 2008; 283(12): 7721 - 7732.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. Llorian, M. Beullens, B. Lesage, E. Nicolaescu, L. Beke, W. Landuyt, J.-M. Ortiz, and M. Bollen
Nucleocytoplasmic Shuttling of the Splicing Factor SIPP1
J. Biol. Chem., November 18, 2005; 280(46): 38862 - 38869.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
C. Lin, L. Yang, J. J. Yang, Y. Huang, and Z.-R. Liu
ATPase/Helicase Activities of p68 RNA Helicase Are Required for Pre-mRNA Splicing but Not for Assembly of the Spliceosome
Mol. Cell. Biol., September 1, 2005; 25(17): 7484 - 7493.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. T. Nasim and R. C. Trembath
A dual-light reporter system to determine the efficiency of protein-protein interactions in mammalian cells
Nucleic Acids Res., April 11, 2005; 33(7): e66 - e66.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
H. H. A. G. M. van der Putten, R. M. Bertina, H. L. Vos, N. von Ahsen, and M. Oellerich
Is the prothrombin 19911A>G polymorphism a functional noncoding variant?
Blood, April 1, 2005; 105(7): 2995 - 2996.
[Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
J. H. Trembley, S. Tatsumi, E. Sakashita, P. Loyer, C. A. Slaughter, H. Suzuki, H. Endo, V. J. Kidd, and A. Mayeda
Activation of Pre-mRNA Splicing by Human RNPS1 Is Regulated by CK2 Phosphorylation
Mol. Cell. Biol., February 15, 2005; 25(4): 1446 - 1457.
[Abstract] [Full Text] [PDF]

Home page
 BloodHome page
N. von Ahsen and M. Oellerich
The intronic prothrombin 19911A>G polymorphism influences splicing efficiency and modulates effects of the 20210G>A polymorphism on mRNA amount and expression in a stable reporter gene assay system
Blood, January 15, 2004; 103(2): 586 - 593.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.