Part:BBa_K4789006
pcDNA-pre-miR-22
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-22 is an oncomiR that is dyregulated in several female cancers, including cervical cancer, breast cancer and ovarian cancer [1]. Our team focus on the cervical cancer development. Previous research have discovered miR-22 are significantly downregulated in cervical cancer1[2-3].A growing number of studies highlights the role of miR-22 in cervical cancer drug resistance development [4]. 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[5]. Pre-mir-22 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-miR22 sequence into the basic part pcDNA 3.0 plasmid. Then the miR-22 could be overexpressed in the Hela cell.
Figure 1 miR-22 and LncRNA binding site
Reference
[1]Nejati Kazem,Alivand MohammadReza,Arabzadeh AmirAhmad,MicroRNA-22 in female malignancies: Focusing on breast, cervical, and ovarian cancers.[J] .Pathol Res Pract, 2021, 223: 153452.
[2]Konishi Hiromi,Hayashi Masami,Taniguchi Kohei et al. The therapeutic potential of exosomal miR-22 for cervical cancer radiotherapy.[J] .Cancer Biol Ther, 2020, 21: 1128-1135.
[3]Zhang Lu,Chen Bin,Ding Ding,Decreased microRNA-22 is associated with poor prognosis in cervical cancer.[J] .Int J Clin Exp Pathol, 2017, 10: 9515-9520.
[4] Nakamura Mayumi,Hayashi Masami,Konishi Hiromi et al. MicroRNA-22 enhances radiosensitivity in cervical cancer cell lines via direct inhibition of c-Myc binding protein, and the subsequent reduction in hTERT expression.[J] .Oncol Lett, 2020, 19: 2213-2222.
[5] 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.
[6] http://www.mircode.org/
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 952
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Illegal PstI site found at 2313 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 952
Illegal NheI site found at 895
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Illegal SpeI site found at 935
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Illegal NotI site found at 978 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 952
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Illegal XhoI site found at 985 - 23INCOMPATIBLE 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 - 25INCOMPATIBLE 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
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Illegal NgoMIV site found at 3047 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 882
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