Regulatory

Part:BBa_K4829005

Designed by: Modernatx   Group: iGEM23_IISc-Bengaluru   (2023-10-04)


3' UTR patented by Modernatx

This 3'UTR was derived from the Beta Globin UTRs, for expression of a non-human protein in a human cell. Please use ONLY for research(academic) purposes. It is illegal to use for other purposes. More information may be available on the US patent page US10925935B2.


Usage and Biology

3' UTRs: Untranslated Regions at the mRNA's End

The 3' UTR, or 3' Untranslated Region, is a crucial segment located at the end of a messenger RNA (mRNA) molecule. While it does not encode proteins, the 3' UTR plays a vital role in regulating gene expression and post-transcriptional processes. Here's why 3' UTRs are important:

  • Polyadenylation and Stability: The 3' UTR contains the polyadenylation signal, which marks the end of the mRNA molecule and is essential for the addition of a poly-A tail. This poly-A tail stabilizes the mRNA, preventing its degradation and ensuring its longevity in the cell. Without a properly structured 3' UTR, mRNA molecules may be rapidly degraded.
  • Regulatory Elements: Like the 5' UTR, the 3' UTR also houses regulatory elements. These include binding sites for various proteins, RNA-binding proteins, and microRNAs. These elements can influence mRNA stability, translation efficiency, and subcellular localization. MicroRNAs, in particular, can bind to complementary sequences in the 3' UTR, leading to mRNA degradation or translational repression.
  • Alternative Polyadenylation: The 3' UTR can undergo alternative polyadenylation, resulting in mRNA isoforms with different 3' UTR lengths. This can have a profound impact on gene regulation, as the length and composition of the 3' UTR can affect how the mRNA is regulated and processed.

Rationale:

  • The use of the β-globin gene's 3' UTR is attributed to its role in mRNA stability. Such stability is vital for the effective translation of the target protein once the mRNA is inside human cells. Ensuring mRNA remains intact and functional for a sufficient duration inside the body is crucial to stimulate an immune response.
  • Stability regulatory elements, especially from globin genes, form RNA-protein complexes that stabilize the mRNA. Given the importance of mRNA stability for vaccine efficacy, this design choice is aligned with the objective of producing a robust and long-lasting immune response.

In academia, where using patented sequences is permitted, as long as the correct acknowledgement is given, one can use these sequences as seen in our BBa K4829003. This is done when the requirement to optimise or profit out of the mRNA is not required. We would like to remind all users, once again, that using these sequences without acknowledgement or using these sequences for making a profit, is STRICTLY PROHIBITED. Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Functional Parameters

We have used this part in BBa K4829003, and we noticed expression in Hela cells, 6hrs post-transfection.

  • hela-blot.png
 Figure 1. Expression of the protein noted in the supernatant 6 hours post transfection ]]
[edit]
Categories
Parameters
None