Non-coding RNAs: regulators of disease
14 important questions on Non-coding RNAs: regulators of disease
What is miRNA function?
Binding of miRNA to target sequence:
mRNA degradation/repression
decrease in protein expression
what can a DNA trancription lead to?
rRNA (ribosomal)
mRNA
tRNA (transfer)
How do you detect miRNAs?
by PCR or microarray methodology
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Biological roles are studied through gain- or loss-of-function of a particular miRNA. Name two examples:
In vitro techniques (shRNA and siRNA)
Knockout mice
How can you predict targets of miRNAs?
using online databases
How do you validate tagerts of miRNA?
using genome-wide transcriptional profiling
Small RNAs have roles in virtually all developmental processes, including:
stem cell and germline maintenance,
development and differentiation,
transcriptional and post-transcriptional gene silencing
subcellular localization
Loss of Dicer in mouse models leads to embryonic lethality
What are the Important roles for miRNA’s in Immunology and Cancer?
miRNAs can act as tumor suppressors/oncogenes
Viral defense against HIV: degradation of viral RNA in human T-cells
Important for development, differentiation, survival and function of immune cells
What is the goal ofmiRNAs control the expression of TLR signalling components?
switching off the pathway to prevent over-amplification of this signal
Interfering RNA’s in Innate immunity, miR-155:
Pro-inflammatory
Targets anti-inflammatory pathways/molecules (repression of SOCS1/SHIP1)
Needed for optimal functioning of Dendritic Cells and T helper 1 cells
Interfering RNA’s in Innate immunity, miR-146:
Negative feed-back: downregulation TLR and cytokine signaling
Inhibits T cell activation
Differential expression in dendritic cells subsets
High expression in Langerhans Cells (to prevent inappropriate activation by commensal bacteria on the skin)
MicroRNAs (miRNAs) as potential diagnostic biomarkers in cancer . Name 5
1) utlizing a signature of altered miRNA expression to differentiate cancer tissue from normal tissue.
2) use of miRNA based classifier to indentify tissue of origin for cancers of unknown primaries.
3) distinguishing tumor subtypes using a panel of miRNas that show differential expression within one cancer type
4) study SNPs in the miRNA genes, miRNA binding sites in the target mRNA genes or in the miRNA processing/machinery pathway genes to predict cancer predisposition
5) profiling circulating blood or tumor derived exosomal miRNAs, surpassing the invasive procedures to aid in early detection of cancers
Use of RNA interference in science:
Genetically manipulate mammalian cells
Specifically silence expression of certain genes
Gain insight into function of proteins and biological pathways of interest
Development of treatments for cancer, AIDS etc.
shRNA (short-hairpin RNA):
Short-hairpin RNA resulting from transfected or transduced vector
Needs processing by Dicer
Can be virally delivered, very efficient method
Renewable and potentially inducible
Drawbacks:
Aspecific targeting
Aspecific effects of viral transduction
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