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Use of minigene systems to dissect alternative splicing elements.
Methods. 2005 Dec; 37(4):331-40.M

Abstract

Pre-mRNA splicing is an essential step for gene expression in higher eukaryotes. The splicing efficiency of individual exons is determined by multiple features involving gene architecture, a variety of cis-acting elements within the exons and flanking introns, and interactions with components of the basal splicing machinery (called the spliceosome) and auxiliary regulatory factors which transiently co-assemble with the spliceosome. Both alternative and constitutive exons are recognized by multiple weak protein:RNA interactions and different exons differ in the interactions which are determinative for exon usage. Alternative exons are often regulated according to cell-specific patterns and regulation is mediated by specific sets of cis-acting elements and trans-acting factors. Transient expression of minigenes is a commonly used in vivo assay to identify the intrinsic features of a gene that control exon usage, identify specific cis-acting elements that control usage of constitutive and alternative exons, identify cis-acting elements that control cell-specific usage of alternative exons, and once regulatory elements have been identified, to identify the trans-acting factors that bind to these elements and modulate splicing. This chapter describes approaches and strategies for using minigenes to define the cis-acting elements that determine splice site usage and to identify and characterize the trans-acting factors that bind to these elements and regulate alternative splicing.

Authors+Show Affiliations

Department of Pathology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA. tcooper@bcm.edu

Pub Type(s)

Journal Article

Language

eng

PubMed ID

16314262

Citation

Cooper, Thomas A.. "Use of Minigene Systems to Dissect Alternative Splicing Elements." Methods (San Diego, Calif.), vol. 37, no. 4, 2005, pp. 331-40.
Cooper TA. Use of minigene systems to dissect alternative splicing elements. Methods. 2005;37(4):331-40.
Cooper, T. A. (2005). Use of minigene systems to dissect alternative splicing elements. Methods (San Diego, Calif.), 37(4), 331-40.
Cooper TA. Use of Minigene Systems to Dissect Alternative Splicing Elements. Methods. 2005;37(4):331-40. PubMed PMID: 16314262.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Use of minigene systems to dissect alternative splicing elements. A1 - Cooper,Thomas A, PY - 2005/07/05/received PY - 2005/07/05/accepted PY - 2005/11/30/pubmed PY - 2006/2/4/medline PY - 2005/11/30/entrez SP - 331 EP - 40 JF - Methods (San Diego, Calif.) JO - Methods VL - 37 IS - 4 N2 - Pre-mRNA splicing is an essential step for gene expression in higher eukaryotes. The splicing efficiency of individual exons is determined by multiple features involving gene architecture, a variety of cis-acting elements within the exons and flanking introns, and interactions with components of the basal splicing machinery (called the spliceosome) and auxiliary regulatory factors which transiently co-assemble with the spliceosome. Both alternative and constitutive exons are recognized by multiple weak protein:RNA interactions and different exons differ in the interactions which are determinative for exon usage. Alternative exons are often regulated according to cell-specific patterns and regulation is mediated by specific sets of cis-acting elements and trans-acting factors. Transient expression of minigenes is a commonly used in vivo assay to identify the intrinsic features of a gene that control exon usage, identify specific cis-acting elements that control usage of constitutive and alternative exons, identify cis-acting elements that control cell-specific usage of alternative exons, and once regulatory elements have been identified, to identify the trans-acting factors that bind to these elements and modulate splicing. This chapter describes approaches and strategies for using minigenes to define the cis-acting elements that determine splice site usage and to identify and characterize the trans-acting factors that bind to these elements and regulate alternative splicing. SN - 1046-2023 UR - https://www.unboundmedicine.com/medline/citation/16314262/Use_of_minigene_systems_to_dissect_alternative_splicing_elements_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1046-2023(05)00173-8 DB - PRIME DP - Unbound Medicine ER -