One Of The New Method Of Discovering Alternative Splicing Isoforms

Gene is transcribed into pre-mRNAs.The pre-mRNAs include intronic and exonic sequences,and undergo various post-transcriptional modifications. One of these key processes is pre-mRNA splicing: a mechanism that removes intronic sequences and joins their surrounding exons. Splicing can be either constitutive or alternative. When a pre-mRNA undergoes constitutive splicing the immature transcript is processed in the same way so that only one type of mRNA is produced and only one protein is generated from a the gene. Through alternative splicing,the pre-mRNA can be processed in different ways to produce various mRNAs,which give rise to a variety of proteins that differ in their peptide sequence,thus, might show different chemical and biological properties.Traditionally, the alternative splicing in a given condition has been analyzed on a gene-by-gene basis using RT-PCR-based techniques. At the single-gene level ,theses techniques does not show the real implications of splicing alterations in cancer. For this reason,splicing specific microarrays have been developed recently. An important limitation is that information about alternative-splicing events in public databases is still fragmented. Microarrays designed on the basis of known transcripts will probably not detect unknown or abnormal splicing variants which is especially important in the diagnosis, prognosis, and treatment of disease, such as in cancer, which can produce several uncharacterized splicing events. So we throw our sight on this point. To find a procedure that is a supplement way to upload as many uncharacterized splicing events of disease state as possible to the databases, and that must be cost effective, sensitive and specific, we developed single gene specific primer pool (GSPP) method.