Difference between revisions of "Part:BBa K1993002"

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Chemokine receptors are receptors found on the surface of certain cells that interact with chemokines. They have a 7-transmembrane (7-TM) structure and couple to G-protein for signal transduction within a cell. [1] (Figure 1) Following interaction with their specific chemokine ligands, chemokine receptors trigger a flux intracellular calcium (Ca2+) ions, initiate chemotaxis and guide the cell to a desired location. (Figure 2)
Chemokine receptors are found on the surface of certain cells that interact with chemokines. They have a 7-transmembrane (7TM) structure and couples to G-protein for signal transduction within a cell[1] (Figure 1). Following interaction with their specific chemokine ligands, chemokine receptors trigger a flux inintracellular calcium (Ca2+) ions which initiate the onset of chemotaxis that traffics the cell to a desired location (Figure 2).
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'''Figure 1. typical structure of a chemokine receptor.'''
 
'''Figure 1. typical structure of a chemokine receptor.'''
  
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<img src="https://static.igem.org/mediawiki/2016/0/02/T--SYSU-MEDICINE--interaction.png" style="width:800px"  ></a>
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'''Figure 2. the mechanism of interaction between chemokine and chemokine receptor.'''
 
'''Figure 2. the mechanism of interaction between chemokine and chemokine receptor.'''
  
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Under the circumstances of inflammation, various kinds of cytokines, including chemokines, are released by the lesions. Guided by the chemokines, cells expressing chemokine receptors move towards the lesions where they can function better.[2] What’s more, different diseases would release different pools of chemokines, which would recruit different effector cells. [https://static.igem.org/mediawiki/2016/2/2d/T--SYSU-MEDICINE--project-diseasse-table-chemokine.pdf See our disease table-chemokine]
  
Under the circumstance of inflammation, various kinds of cytokines, including chemokines, are released by the lesions. Guided by the chemokines, cells expressing chemokine receptors move towards the lesions where they can function better. [2] What’s more, different diseases would release different pool of chemokines, which would recruit different effector cells.[https://static.igem.org/mediawiki/2016/2/2d/T--SYSU-MEDICINE--project-diseasse-table-chemokine.pdf See our disease talbe-chemokine]
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Based on the chemotaxis theory, in order to enhance the homing ability of our marvelous mesenchymal stem cells (MSCs) due to lack of enough chemokine receptors on their cell surface, we, SYSU-MEDICINE, had constructed a series of chemokine receptors that corresponding to different inflammatory diseases as far as possible. Among which, CXCL6 is a significant chemokine (CXCR1 is its chemokine receptor) in Inflammatory Bowel Disease(IBD). [https://static.igem.org/mediawiki/2016/2/2d/T--SYSU-MEDICINE--project-diseasse-table-chemokine.pdf See our disease table-chemokine]
  
 
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We acquired this gene from peripheral mononuclear blood cells (PMBCs) and purified it. (Figure 3) Then we constructed it under the control of EF-by Gateway technology. (Figure 4)
Based on the chemotaxis theory, in order to enhance the homing ability of our marvelous mesenchymal stem cells(MSCs) since their lack of enough chemokine receptors on their cell surface,  we, SYSU-MEDICINE, had constructed a series of chemokine receptors that cover different inflammatory diseases as far as possible. Among which, CXCR1 is a significant chemokine receptor in Arthritis.[https://static.igem.org/mediawiki/2016/2/2d/T--SYSU-MEDICINE--project-diseasse-table-chemokine.pdf See our disease talbe-chemokine]
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We got the gene from peripheral mononuclear blood cells (PMBCs) and purified it (Figure 3). Then we constructed it under the control of EF1-alpha by using Gateway technology.(Figure 4)
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<img src="https://static.igem.org/mediawiki/2016/d/d7/T--SYSU-MEDICINE--CCR7-PCR.png" style="width:100px"  ></a>
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<img src="https://static.igem.org/mediawiki/2016/3/37/T--SYSU-MEDICINE--CXCR1.jpeg" style="width:150px"  ></a>
  
 
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'''Figure 3. Purified of CXCR1.'''
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'''Figure 3 purification of gene CXCR1'''
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<img src="https://static.igem.org/mediawiki/2016/4/47/T--SYSU-MEDICINE--EF1-a-CCR7.png" style="width:200px"  ></a>
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<img src="https://static.igem.org/mediawiki/2016/2/27/T--SYSU-MEDICINE--1.2.3.png" style="width:200px"  ></a>
  
 
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'''Figure 4. EF1-a CXCR1.'''
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'''Figure 4 Construction of expression vector of CXCR1'''
  
We introduced that plasmid into MSCs and tested the expression of CXCR1 in MSCs on the protein level (Figure 5)
 
  
 
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Then, we tested the chemotaxis of engineered MSCs by conducting Transwell assay against CXCL6. To our excitement, our engineered MSCs had improved their homing ability with chemokine receptor CXCR1(Figure 6).
 
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'''Figure 5. Western Blot Of CXCR1.'''
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Then, we tested the chemotaxis of engineered MSCs by conducting Transwell assay against CXCL3. To our excitement, our engineered MSCs had improved their homing ability with chemokine receptor CXCR1(Figure 6).
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'''Figure 6. transwell assay of CXCR1.'''
 
'''Figure 6. transwell assay of CXCR1.'''
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[2] Griffith J W, Sokol C L, Luster A D. Chemokines and chemokine receptors: positioning cells for host defense and immunity.[J]. Annual Review of Immunology, 2014, 32(1):659-702.
 
[2] Griffith J W, Sokol C L, Luster A D. Chemokines and chemokine receptors: positioning cells for host defense and immunity.[J]. Annual Review of Immunology, 2014, 32(1):659-702.
 
 
  
 
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Revision as of 06:55, 16 October 2016


CXCR1

Chemokine receptors are receptors found on the surface of certain cells that interact with chemokines. They have a 7-transmembrane (7-TM) structure and couple to G-protein for signal transduction within a cell. [1] (Figure 1) Following interaction with their specific chemokine ligands, chemokine receptors trigger a flux intracellular calcium (Ca2+) ions, initiate chemotaxis and guide the cell to a desired location. (Figure 2)

Figure 1. typical structure of a chemokine receptor.

Figure 2. the mechanism of interaction between chemokine and chemokine receptor.

Under the circumstances of inflammation, various kinds of cytokines, including chemokines, are released by the lesions. Guided by the chemokines, cells expressing chemokine receptors move towards the lesions where they can function better.[2] What’s more, different diseases would release different pools of chemokines, which would recruit different effector cells. See our disease table-chemokine

Based on the chemotaxis theory, in order to enhance the homing ability of our marvelous mesenchymal stem cells (MSCs) due to lack of enough chemokine receptors on their cell surface, we, SYSU-MEDICINE, had constructed a series of chemokine receptors that corresponding to different inflammatory diseases as far as possible. Among which, CXCL6 is a significant chemokine (CXCR1 is its chemokine receptor) in Inflammatory Bowel Disease(IBD). See our disease table-chemokine

We acquired this gene from peripheral mononuclear blood cells (PMBCs) and purified it. (Figure 3) Then we constructed it under the control of EF-1α by Gateway technology. (Figure 4)


Figure 3 purification of gene CXCR1


Figure 4 Construction of expression vector of CXCR1


Then, we tested the chemotaxis of engineered MSCs by conducting Transwell assay against CXCL6. To our excitement, our engineered MSCs had improved their homing ability with chemokine receptor CXCR1(Figure 6).

Figure 6. transwell assay of CXCR1.

References

[1]Allen, Samantha J.; Crown, Susan E.; Handel, Tracy M. (2007-01-01). "Chemokine: receptor structure, interactions, and antagonism". Annual Review of Immunology. 25: 787–820.

[2] Griffith J W, Sokol C L, Luster A D. Chemokines and chemokine receptors: positioning cells for host defense and immunity.[J]. Annual Review of Immunology, 2014, 32(1):659-702.

Sequence and Features


Assembly Compatibility:
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    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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1111
    Illegal SapI site found at 1017