Here, we report findings attributing to impairments in the splicing process such a regulatory role.
Many play fundamental roles in the splicing and translation of protein-coding RNAs, whereas others play regulatory roles.
Mutations in the splicing machinery underlie most cases of a bone marrow cancer called myelodisplastic syndrome, for example.
As polyadenylation is known to precede splicing2,3, the possibility exists that every transcript possesses sequences near the 3′ end that are directly involved in the splicing process.
The SV40 system has been of particular value in elucidating certain genetic elements which are critically involved in the splicing process17 21.
Vitulo, N. et al. A deep survey of alternative splicing in grape reveals changes in the splicing machinery related to tissue, stress condition and genotype.
Kim, S. S. et al. Loss-of-function mutations in the splicing factor ZRSR2 are common in blastic plasmacytoid dendritic cell neoplasm and have male predominance.
They also hope to identify other genes and proteins involved in the splicing process they discovered, which could also make good drug targets.
We decided to investigate a NLS in Trypanosoma cruzi by selecting p14, a small RNA recognition motif (RRM) containing protein involved in the splicing process in the nucleus.
Regarding that the limitation of co-amplification PCR may not fully reflect alternative splicing scenario, we further conducted qRT-PCR assay using isoform-specific primers to confirm there are indeed reciprocal changes in the splicing variant (Supplementary Fig. 17).
Due to limitation of PCR reaction in co-amplification, we further performed RT-qPCR of individual splicing variants from each of the selected gene, and found that there are reciprocal changes in the splicing variant levels (Supplementary Fig. 17).
