Premium
Monitoring changes in alternative precursor messenger RNA splicing in multiple gene transcripts
Author(s) -
Simpson Craig G.,
Fuller John,
Maronova Monika,
Kalyna Maria,
Davidson Diane,
McNicol Jim,
Barta Andrea,
Brown John W. S.
Publication year - 2008
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2007.03392.x
Subject(s) - biology , alternative splicing , gene , rna splicing , genetics , messenger rna , arabidopsis , rna , gene expression , computational biology , precursor mrna , microbiology and biotechnology , mutant
Summary Alternative splicing (AS) increases the proteomic and functional capacity of genomes through the generation of alternative mRNA transcripts from the same gene. AS is now estimated to occur in a third of Arabidopsis and rice genes, and includes genes involved in the control of growth and development, responses to stress and signalling. Regulation of AS reflects the interactions between positive and negative cis sequences in the precursor messenger RNA and a range of trans ‐acting factors. The levels and activities of these factors differ in different cells and growth conditions. To identify changes in AS in multiple genes simultaneously, we have established a reproducible RT‐PCR panel that can analyse 96 alternative splicing events and accurately measure the ratio of alternatively spliced products. This procedure detected statistically significant changes in AS in different plant organs, in plants grown under different light and day‐length conditions, and in plants overexpressing splicing factors. The system provides a convenient, medium‐throughput means of monitoring changes in AS in multiple genes. It can readily be applied to much larger or targeted sets of gene transcripts to generate information on the significance and regulation of AS in plant growth and development, specific processes and responses to external stimuli.