RT Journal
A1 Thill, Gilbert
A1 Castelli, Vanina
A1 Pallud, Sophie
A1 Salanoubat, Marcel
A1 Wincker, Patrick
A1 de la Grange, Pierre
A1 Auboeuf, Didier
A1 Schächter, Vincent
A1 Weissenbach, Jean
T1 ASEtrap: A biological method for speeding up the exploration of spliceomes
JF Genome Research 
JO Genome Research 
YR 2006 
FD June 01 
VO 16 
IS 6 
SP 776 
OP 786 
DO 10.1101/gr.5063306 
UL http://genome.cshlp.org/content/16/6/776.abstract 
AB Alternative splicing (AS) of pre-messenger RNA is a major mechanism for generating protein diversity from a limited number of genes in higher eukaryotes, and it constitutes a central mode of genetic regulation. Thus, efficient methods are needed to systematically identify new AS events at a genomic scale across different tissues, stages of development, and physiological or pathological conditions in order to better understand gene expression. To fulfill this goal, we have designed the ASEtrap, which is a cloning procedure for producing AS libraries that is based on a single-stranded trap consisting of an ssDNA-binding protein. In this paper, we have applied our approach to the construction of an AS library and a Control library from human placenta. By analyzing 9226 and 9999 sequences of the AS and Control libraries, respectively, we show that internal AS events (events that can be identified by the sole resources provided by either the AS or the Control library) and the discovery rate of new AS events measured at early stages of sequencing were nine to 10 times higher in the former than in the latter. Moreover, by performing a search for new AS events within a group of 162 known drug target genes, we identified six new events in six genes, and we observed that they all were discovered exclusively through the AS library. Thus, it appears that ASEtrap has the potential to greatly facilitate the determination of the total complement of splice variants expressed in human, as well as other organisms.