Composite

Part:BBa_K4789007

Designed by: Fangyuan Duan   Group: iGEM23_YiYe-China   (2023-09-23)


pcDNA-pre-miR-145

Usage and Biology

MicroRNAs (miRNAs), a novelty-defined class of regulatory genes, have revolutionized principles of classical bimolecular. These RNAs regulate the expression of a gene through inhibition of translational initiation or targeting mRNAs for degradation. MiR-145 has the diagnose value in patients with cervical cancer[1].Our team also focus on the cervical cancer screening. Previous researches have discovered miR-145 could inhibit cervical cancer progression through targeting WNT2B by Wnt/β-catenin pathway[2] or miR-145/RCAN3 axis[3] and so on. LncRNAs work as ‘miRNA sponges’ through binding to specific miRNAs. Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), long non-coding RNA, has been reported to be upregulated in cervical cancer tissues and promote cancer cell invasion and metastasis[4]. Pre-mir-145 could bind to the sequence of MALAT1 according to the prediction database miRcode(Figure 1)[5]. This mechanism gives rise to our idea about fusing a sponge RNA based on the sequences of lncRNA with binding sites complementary to the sequence of miRNA into a plasmid contained the reporter gene. In our study, we cloned the pre-miR145 sequence into the basic part pcDNA 3.0 plasmid. Then the miR22 could overexpressed in the Hela cell. 

            145-base.png
                  Figure 1   miR-145and LncRNA binding site

Reference

[1]Yu Furong,Liu Jie,Dong Weilei et al. The diagnostic value of miR-145 and miR-205 in patients with cervical cancer.[J] .Am J Transl Res, 2021, 13: 1825-1832.

[2 ]Li Qi,Yu Xiaohong,Yang Liangliang,MiR-145 inhibits cervical cancer progression and metastasis by targeting WNT2B by Wnt/β-catenin pathway.[J] .Int J Clin Exp Pathol, 2019, 12: 3740-3751.

[3]Zhang Xian-Yu,Ma Huan,Li Jing et al. Functional implications of miR-145/RCAN3 axis in the progression of cervical cancer.[J] .Reprod Biol, 2020, 20: 140-146.

[4]Han Xiting,Wang Qian,Wang Yan et al. Long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1/microRNA-202-3p/periostin axis modulates invasion and epithelial-mesenchymal transition in human cervical cancer.[J] .J Cell Physiol, 2019, 234: 14170-14180.

[5] http://www.mircode.org/


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 952
    Illegal XbaI site found at 991
    Illegal SpeI site found at 249
    Illegal SpeI site found at 935
    Illegal PstI site found at 957
    Illegal PstI site found at 2313
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 952
    Illegal NheI site found at 895
    Illegal SpeI site found at 249
    Illegal SpeI site found at 935
    Illegal PstI site found at 957
    Illegal PstI site found at 2313
    Illegal NotI site found at 978
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 952
    Illegal BglII site found at 12
    Illegal BamHI site found at 929
    Illegal XhoI site found at 985
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 952
    Illegal XbaI site found at 991
    Illegal SpeI site found at 249
    Illegal SpeI site found at 935
    Illegal PstI site found at 957
    Illegal PstI site found at 2313
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 952
    Illegal XbaI site found at 991
    Illegal SpeI site found at 249
    Illegal SpeI site found at 935
    Illegal PstI site found at 957
    Illegal PstI site found at 2313
    Illegal NgoMIV site found at 1423
    Illegal NgoMIV site found at 2764
    Illegal NgoMIV site found at 3047
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 882
    Illegal BsaI.rc site found at 4572
    Illegal SapI site found at 3492
    Illegal SapI.rc site found at 2613
    Illegal SapI.rc site found at 2823


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