Difference between revisions of "Part:BBa K4789000"
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Given the fact that lncRNA MALAT1 could act as miRNA sponge, we designed the sequence that could bind to miR-22 based on MALAT1 sequence.
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__NOTOC__
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<partinfo>BBa_K4789000 short</partinfo>
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===Usage and Biology===
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LncRNAs, that share miRNA response elements (MREs) with coding RNAs, contained similar miRNA target sequences and then prevented miRNAs from acting on mRNAs. These lncRNAs, known as competing endogenous RNAs (ceRNAs), served as ‘sponges’ for miRNAs, inhibited the corresponding miRNAs and contributed to enhanced translations of their target mRNAs [1](Fig 1). Currently, Reports have shown that the lncRNA MALAT1 can act as a "sponge" for miR-202-3p, thereby regulating the invasion and epithelial-mesenchymal transition of cervical cancer cells [2]. Additionally, it has been reported that MALAT1 can also act as a "sponge" for miR-429, thereby regulating the migration, apoptosis, and other metabolic activities of cervical cancer cells [3-4].
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                        https://static.igem.wiki/teams/4789/wiki/sponge.png
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                        Figure 1 Molecular mechanisms of lncRNA-miRNA axes.
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  1. MiRNAs regulate gene expression by binding to 3’ UTR elements in mRNA and degrading them.
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  2.lncRNAs have been thought to interact with miRNAs as “sponges” or competing  endogenous RNAs (ceRNA), attenuating the
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  repression of mRNAs by miRNA
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===Design===
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Mir-22 was associated with the metastatic process, and showed potential as biomarker for prediction of metastasis and prognosis of cervical cancer[5] (Fig 2). Mir-22 contained the binding site with lncRNA MALAT1. Given the fact that lncRNA MALAT1 could act as miRNA sponge, we designed the MALAT1 sequence that could bind to miR-22.
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            https://static.igem.wiki/teams/4789/wiki/mir-22-hierarchical-clustering.png
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    Figure 2 Hierarchical clustering of miR-22. NC, normal squamous epithelium
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    SqC-NM, squamous cell carcinoma without metastasis; SqC-M, squamous cell carcinoma with metastasis.
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    The color indicates the signal value, from red (upregulation) to green (downregulation).
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===Characterization===
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The miR-22-sponge sequence was confirmed by Sanger sequencing.  
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                  https://static.igem.wiki/teams/4789/wiki/mir-22-sanger.png
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                    Figure 3 miR-22-sponge validated by Sanger sequencing
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===Reference ===
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1.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.
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2.Shen Fujin,Zheng Hongyun,Zhou Limei et al. Overexpression of MALAT1 contributes to cervical cancer progression by acting as a sponge of miR-429.[J] .J Cell Physiol, 2019, 234: 11219-11226.
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3.Aftab Mehreen,Poojary Satish S,Seshan Vaishnavi et al. Urine miRNA signature as a potential non-invasive diagnostic and prognostic biomarker in cervical cancer.[J] .Sci Rep, 2021, 11: 10323.
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4.Chen Mingming,Ma Zhao,Wu Xiaotian et al. A molecular beacon-based approach for live-cell imaging of RNA transcripts with minimal target engineering at the single-molecule level.[J] .Sci Rep, 2017, 7: 1550.
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5.Kwon Ah-Young,Jeong Ju-Yeon,Park Hyun et al. miR-22-3p and miR-30e-5p Are Associated with Prognosis in Cervical Squamous Cell Carcinoma.[J] .Int J Mol Sci, 2022, 23: undefined.
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<!-- Add more about the biology of this part here
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===Usage and Biology===
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<!-- -->
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<!-- -->
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K4789000 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K4789001 parameters</partinfo>
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Latest revision as of 07:42, 6 October 2023


miR-22-sponge

Usage and Biology

LncRNAs, that share miRNA response elements (MREs) with coding RNAs, contained similar miRNA target sequences and then prevented miRNAs from acting on mRNAs. These lncRNAs, known as competing endogenous RNAs (ceRNAs), served as ‘sponges’ for miRNAs, inhibited the corresponding miRNAs and contributed to enhanced translations of their target mRNAs [1](Fig 1). Currently, Reports have shown that the lncRNA MALAT1 can act as a "sponge" for miR-202-3p, thereby regulating the invasion and epithelial-mesenchymal transition of cervical cancer cells [2]. Additionally, it has been reported that MALAT1 can also act as a "sponge" for miR-429, thereby regulating the migration, apoptosis, and other metabolic activities of cervical cancer cells [3-4]. 

                        sponge.png
                        Figure 1 Molecular mechanisms of lncRNA-miRNA axes.
 1. MiRNAs regulate gene expression by binding to 3’ UTR elements in mRNA and degrading them.
 2.lncRNAs have been thought to interact with miRNAs as “sponges” or competing  endogenous RNAs (ceRNA), attenuating the 
 repression of mRNAs by miRNA

Design

Mir-22 was associated with the metastatic process, and showed potential as biomarker for prediction of metastasis and prognosis of cervical cancer[5] (Fig 2). Mir-22 contained the binding site with lncRNA MALAT1. Given the fact that lncRNA MALAT1 could act as miRNA sponge, we designed the MALAT1 sequence that could bind to miR-22. 

           mir-22-hierarchical-clustering.png
    Figure 2 Hierarchical clustering of miR-22. NC, normal squamous epithelium
    SqC-NM, squamous cell carcinoma without metastasis; SqC-M, squamous cell carcinoma with metastasis. 
    The color indicates the signal value, from red (upregulation) to green (downregulation).

Characterization

The miR-22-sponge sequence was confirmed by Sanger sequencing. 


                  mir-22-sanger.png
        
                   Figure 3 miR-22-sponge validated by Sanger sequencing

Reference

1.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.

2.Shen Fujin,Zheng Hongyun,Zhou Limei et al. Overexpression of MALAT1 contributes to cervical cancer progression by acting as a sponge of miR-429.[J] .J Cell Physiol, 2019, 234: 11219-11226.

3.Aftab Mehreen,Poojary Satish S,Seshan Vaishnavi et al. Urine miRNA signature as a potential non-invasive diagnostic and prognostic biomarker in cervical cancer.[J] .Sci Rep, 2021, 11: 10323.

4.Chen Mingming,Ma Zhao,Wu Xiaotian et al. A molecular beacon-based approach for live-cell imaging of RNA transcripts with minimal target engineering at the single-molecule level.[J] .Sci Rep, 2017, 7: 1550.

5.Kwon Ah-Young,Jeong Ju-Yeon,Park Hyun et al. miR-22-3p and miR-30e-5p Are Associated with Prognosis in Cervical Squamous Cell Carcinoma.[J] .Int J Mol Sci, 2022, 23: undefined.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 127
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 127
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 127
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 127
  • 1000
    COMPATIBLE WITH RFC[1000]