Difference between revisions of "Part:BBa K2957007"

Revision as of 03:12, 22 October 2019

hEF1a, IL-8 (CXCL-8), NeonGreen Tag

Contents: hEF1a, IL-8 (CXCL-8), NeonGreen Tag, SynthpA, with Carb resistance

What is it?: A Composite part; it is a genetic circuit with hEF1a promoter and IL-8 gene allowing for constitutive IL-8 expression

What does it do?: It's responsible for the expression of the chemokine, IL-8 (CXCL8) with Flag tag. It was important for our project in particular because of its involvement in the immune system and is known to create neutrophil swarming.

How to use it?: This part was used for Type IIS Cloning and transfected into HEKs. It also has a NeonGreen tag attached to the IL8 so this allows us to visualize it and determine transfection efficiency. See more on the Parts Overview Page or MIT iGEM 2019 wiki: <a href=https://2019.igem.org/Team:MIT/Parts>."

Characterization

Our team’s best composite part is the hEF1a IL8-NeonGreen construct as we were able to prove it’s functionality in engineering our leader cells, HEK-293T, to secrete a major chemokine of interest to us, IL8, which is involved in inflammatory response and has been shown to lead to neutrophil swarming (secretion test results below). IL8 induces mast cells to produce histamines, which causes openings in the endothelium, which allows neutrophils to move through. We were also able to test the version of this plasmid (BBa_K2957003) with our follower cells, HL60s, in an ibidi chamber (see video on Results page of MIT iGEM 2019 wiki).

Sequence and Features