Team:Heidelberg/Project/miRNA Kit

From 2010.igem.org

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The miTuner was [https://2010.igem.org/3A_Assembly assembled] out of different [https://2010.igem.org/Team:Heidelberg/Parts parts]. Cloning was done following [https://2010.igem.org/Team:Heidelberg/Notebook/Material_Methods standard protocols].
The miTuner was [https://2010.igem.org/3A_Assembly assembled] out of different [https://2010.igem.org/Team:Heidelberg/Parts parts]. Cloning was done following [https://2010.igem.org/Team:Heidelberg/Notebook/Material_Methods standard protocols].
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Since the miTuner was constructed initially for Dual-Luciferase assay, this was the method of choice for tuning quantification:
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Since the miTuner was constructed initially for [https://2010.igem.org/Team:Heidelberg/Notebook/Material_Methods#Dual_Luciferase_Assay Dual Luciferase Assay] assay, this was the method of choice for tuning quantification.
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* [https://2010.igem.org/Team:Heidelberg/Notebook/Material_Methods#Dual_Luciferase_Assay Dual Luciferase Assay]
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==References==
==References==

Revision as of 13:50, 24 October 2010

Synthetic miRNA Kit

Abstract

Introduction

MicroRNAs (miRNAs) are endogenous 22-nt long non-coding RNAs that regulate gene expression post-transcriptionally in a diversity of organisms (Bartel, 2004). Although the understanding of miRNA biological functions is progressing remarkably, the exact mechanisms by which miRNAs regulate gene expression are still not unambiguously defined. However, it is commonly believed that miRNAs trigger target mRNA regulation by binding to 3’UTRs (Chekuleva & Filipowicz, 2009). The discovery of the first miRNA (lin-4) revealed sequence complementarity to multiple conserved sites in the 3’UTR of the lin-14 mRNA (Lee et al., 1993; Wightman et al., 1993). Exact principles of gene expression knockdown mediated by miRNA are still in debate (Eulalio et al., 2008).
Binding site properties have an essential impact on miRNA-mRNA interaction. Rational design of binding sites could thereby help to achieve a narrow range of knockdown. The applicability is still limited by redundant target sites and various miRNA expression patterns within the cells which are hardly to measure. The experimental identification and/or validation of miRNA targets by means of proteomic or transcriptomic studies, reporter gene assays or over-expression/knockdown analysis is necessary to understand the biological relevance of the miRNA-mRNA interaction. Although progressively improving, even the reliability of in silico target prediction tools is still far away from being precise.

Results

Discussion

Methods

The miTuner was assembled out of different parts. Cloning was done following standard protocols.

Since the miTuner was constructed initially for Dual Luciferase Assay assay, this was the method of choice for tuning quantification.

References

Contents