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Part:BBa_K2440009

Designed by: Chushu Zhu   Group: iGEM17_NUDT_CHINA   (2017-10-27)
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CMV promoter ->Flag tag -> hAgo2-> SV40 PA

This is the hAgo2 expressing plasmid.

Usage and Biology

Argonaute 2 gene belongs to the argonaute gene family, which encodes for four characteristic domains: N- terminal, PAZ, Mid and a C-terminal PIWI domain. 2

The PAZ domain is named after proteins PIWI, AGO, and Zwille. It binding with the 3' end of siRNA and miRNA due to recognizes the sequence independent manner. The miRNA binds to mRNAs in the partially-complementary sites and reduces its stability and expression levels. 3

The C-terminal PIWI domain is named after Drosophila PIWI protein. It participates in the cleavage activity of his target. 4

Argonaute proteins associate with small RNAs that guide mRNA degradation, translational repression, or a combination of both. The high sequence specificity of Argonaute 2 gene in RNA interference phenomenon in the protein may make it become a suitable treatment for the diseases with elevated expression level of specific gene such as pancreatic cancer. 5

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 3288
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 717
    Illegal NheI site found at 3425
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1863
    Illegal XhoI site found at 2773
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 3288
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 3288
    Illegal NgoMIV site found at 726
    Illegal AgeI site found at 2561
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1423


Reference

[1] Meister G, Landthaler M, Patkaniowska A, Dorsett Y, Teng G, Tuschl T.(2004 Jul). "Human Argonaute2 mediates RNA cleavage targeted by miRNAs and siRNAs". Mol Cell. 23;15(2):185-97. PMID: 15260970 DOI: 10.1016/j.molcel.2004.07.007

[2] Hutvagner G, Simard MJ. (2008). "Argonaute proteins: key players in RNA silencing". Nature Reviews Molecular Cell Biology. 9 (1): 22–32. DOI: 10.1038/nrm2321

[3] Tang G. (Feb 2005). "siRNA and miRNA: an insight into RISCs.". Trends in Biochemical Sciences. 30 (2): 106–14. PMID 15691656. DOI: 10.1016/j.tibs.2004.12.007.

[4] Meister G, Landthaler M, Patkaniowska A, Dorsett Y, Teng G, Tuschl T. (Jul 2004). "Human Argonaute2 Mediates RNA Cleavage Targeted by miRNAs and siRNAs". Molecular Cell. 15 (2): 185–197. PMID 15260970. DOI: 10.1016/j.molcel.2004.07.007.

[5] Ronemus M, Vaughn MW, Martienssen RA. (2006 Jul). "MicroRNA-targeted and small interfering RNA-mediated mRNA degradation is regulated by argonaute, dicer, and RNA-dependent RNA polymerase in Arabidopsis". Plant Cell. 18(7):1559-74. Epub 2006 Jun 23. PMID: 16798886 PMCID: PMC1488920 DOI: 10.1105/tpc.106.042127

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