Recently, repair of defective splicing by small nuclear RNAs (SnRNAs
), as a therapeutic agent, was proposed.
In the vertebrate nucleus, splicing is performed by the spliceosome, a ribosome-sized complex that contains mostly protein, but also includes, at various stages of the reaction, five snRNAs
. It has been proposed that the contemporary spliceosome descended from an RNA-only enzyme, and that some of the snRNAs
may still catalyze the transesterification reactions involved in splicing.
Therefore, in the present study, we assessed the abundance pattern of 887 miRNAs, also including 1 snRNA
, in serum of ovarian cancer patients in comparison to healthy controls and investigated their potential to aid diagnosis and therapy monitoring.
miRNA Assay number miR-125a 002198 miR-192 000491 miR-200b 002251 miR-375 000564 miR-30a 000417 miR-107 000443 miR-143 463509 U6 snRNA
GAPDH and U6 snRNA
(Toyobo, Osaka, Japan) levels were measured and used to normalize the relative abundance of mRNA and miRNA, respectively.
We choose the RNU6-6P snRNA
control for normalization, because its expression was not altered by our experimental conditions.
U6 small nuclear RNA (snRNA
) expression was used as the internal control.
The expression of miR-33 was normalized to U6 snRNA
However, the presence of other small RNAs, such as ribosomal fragments, tRNA, snRNA
, and mtRNA was approximately 12% and 17% of mapped sequence reads of low and high RFI libraries respectively.
We also solved the structure of U4/U6.U5 tri-snRNP by cryoEM and revealed the nearly complete organisation of U5 snRNA
and U4/U6 snRNA
and over 30 proteins within this complex, providing crucial insights into the activation mechanism and the active site of the spliceosome.
was used as internal standard to normalize the expression.
For example, Oct-1 activates snRNA
transcription by cooperating with SNAPc , whereas the Oct-1 and Oct-2 heterocomplex represses transcription of inducible nitric oxide synthase (iNOS) .