Difference between revisions of "Part:BBa K1391108:Experience"

Revision as of 22:52, 1 November 2014

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Applications of BBa_K1391108

Lyn is a Tyrosine-protein kinase responsible for phosphorylating the immunoreceptor tyrosine-based activation motif (ITAM) of a B-Cell Receptor (BCR) Complex upon antigen binding. When the BCR binds it's specific antigen the receptor dimerizes and recruits Lyn which phosphorylates the ITAMs of the CD79A and CD79B proteins in the BCR Complex. This allows the receptor to phosphorylate Syk and initiate a downstream signalling cascade. This part is under a constitutive hEF1a promoter for mammalian cells.

B-cell receptors (BCRs) are naturally occurring, transmembrane protein complexes that consist of a membrane-bound antibody (IgM) and some associated proteins (CD79A and CD79B). Given that the variable region of the antibody can be specific for any of a large number of antigens, we designed a B-cell receptor to bind beta-amyloid plaques (a biomolecular hallmark of Alzheimer's disease). Once bound, activated receptors instigate intracellular signalling, which can then be manipulated to diagnose the disease.

User Reviews

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 ===Applications of BBa_K1391108===
-B-cell receptors (BCRs) are multiprotein immune receptors found exclusively on the surface of B cells. The BCR multiprotein complex is centered around a membrane-bound IgM antibody. When the antibody binds to an extracellular antigen, receptors dimerize resulting in the phosphorylation of the intracellular tails of CD79A and CD79B by the tyrosine-protein kinase Lyn. In response, another cofactor, spleen tyrosine kinase (Syk), is recruited to the receptor and phosphorylated, initiating a signalling cascade that results in the proliferation of the activated B cells. This receptor is important in clonal selection of B cells during human immune response.
+Lyn is a Tyrosine-protein kinase responsible for phosphorylating the immunoreceptor tyrosine-based activation motif (ITAM) of a B-Cell Receptor (BCR) Complex upon antigen binding. When the BCR binds it's specific antigen the receptor dimerizes and recruits Lyn which phosphorylates the ITAMs of the CD79A and CD79B proteins in the BCR Complex. This allows the receptor to phosphorylate Syk and initiate a downstream signalling cascade.
+This part is under a constitutive hEF1a promoter for mammalian cells.
-For this project, we engineered a BCR to respond to beta-amyloid plaques, the hallmark of Alzheimer's disease. This task was accomplished by using a beta-amyloid specific variable region [derived from Gantenerumab] in the membrane-bound IgM antibody. Our design was based on that of the Tango system [1], which capitalizes on the interaction between TEV protease (TEVp) and its cleavage site (TCS), an amino acid sequence for which the protease has a high affinity. A TEV cleavage site was used to link a transcriptional activator (Gal4VP16) to the intracellular tails of BCR accessory proteins CD79A and CD79B, and the receptor’s cofactor, Syk, was fused to TEV protease. Thus, when the modified receptor activates upon binding its antigen, beta-amyloid, Syk-TEVp fusion protein is recruited, bringing TEVp in close proximity to its cleavage site. This proximity of TEVp to TCS results in the cleavage of the transcriptional activator from the receptor releasing it to activate downstream gene circuits.
-The engineered BCR we developed binds beta amyloid with high specificity and releases a transcriptional activator upon binding, making it an extremely valuable tool in the detection of Alzheimer’s Disease. More importantly, the IgM antibody that determines what the receptor binds can be easily swapped out as can the transcription factor the receptor releases. This means that the receptor we developed can bind to any molecule that an antibody can be produced against and it can release any transcription factor in response to the binding of the target molecule. This modularity allows this receptor to be generalized to almost any extracellular sensing making it an invaluable part of any synthetic biologists toolkit.
-This plasmid codes for the protein sequence Lyn. Lyn is a protein-tyrosine kinase which most noticeably plays an important role in B-Cell Receptor activation and phosphorylation.
+B-cell receptors (BCRs) are naturally occurring, transmembrane protein complexes that consist of a membrane-bound antibody (IgM) and some associated proteins (CD79A and CD79B). Given that the variable region of the antibody can be specific for any of a large number of antigens, we designed a B-cell receptor to bind beta-amyloid plaques (a biomolecular hallmark of Alzheimer's disease). Once bound, activated receptors instigate intracellular signalling, which can then be manipulated to diagnose the disease.
 ===User Reviews===