Difference between revisions of "Part:BBa K1993001"

Revision as of 14:49, 17 October 2016

CCR7

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 talbe-chemokine]

We transfected the plasmid into MSCs and confirmed whether CCR7 were expressed in MSCs on the protein level(Figure 5)

Finally, we tested the chemotaxis of our engineered MSCs by conducting transwell assay against CCL19. To our excitement, our engineered MSCs had improved their homing ability after acquiring chemokine receptor CCR7. (Figure 6).

Figure 3. Purified of CCR7.

Figure 4. EF1-a CCR7.

We introduced that plasmid into MSCs and tested the expression of CCR7 in MSCs on the protein level (Figure 5)

Figure 5. Western Blot Of CCR7.

Then, we tested the chemotaxis of engineered MSCs by conducting Transwell assay against CCL17. To our excitement, our engineered MSCs had improved their homing ability with chemokine receptor CCR7(Figure 6).

Figure 6. transwell assay of CCR7.

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.doi:10.1146/annurev.immunol.24.021605.090529. ISSN 0732-0582. PMID 17291188.

[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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Illegal BsaI.rc site found at 1111
Illegal SapI site found at 1017

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 <partinfo>BBa_K1993001 short</partinfo>
-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).
+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)
 <html>
@@ Line 20: / Line 20: @@
-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 table-chemokine]
+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 talbe-chemokine]
+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 talbe-chemokine]
-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, CCR7 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 table-chemokine]
+We transfected the plasmid into MSCs and confirmed whether CCR7 were expressed in MSCs on the protein level(Figure 5)
+Finally, we tested the chemotaxis of our engineered MSCs by conducting transwell assay against CCL19. To our excitement, our engineered MSCs had improved their homing ability after acquiring chemokine receptor CCR7. (Figure 6).
-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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 ==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.
+[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.doi:10.1146/annurev.immunol.24.021605.090529. ISSN 0732-0582. PMID 17291188.
 [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.