Mentor(s): Timothy Wilson, Ph.D.

B Cells are a type of lymphocyte whose primary purpose is to generate highly specific antibodies in response to a foreign antigen. The B cell receptor (BCR) signaling pathway regulates B cell activation, and Fc receptor-like proteins may contribute to this pathway’s activation or inhibition. Previous studies have demonstrated that Fc receptor-like protein 1 (FCRL1) is found on B cell plasma membranes, associating with the BCR and enhancing the activation of the B cell. Because it is unknown which region of FCRL1 is responsible for its recruitment to the BCR, the FCRL1 localization pathway is a relevant and important topic in research to further our understanding of B cell activation. In this study, we use stitching PCR to amplify the DNA pertaining to either the transmembrane and cytoplasmic portion or the transmembrane and extracellular portion of FCRL1 to create two constructs which will then be inserted into the genome of A20 mouse B cells using a lentiviral vector. Cells will then be activated and studied under confocal microscopy to determine which region mediates FCRL1 localization to the BCR. Early results include successful PCR amplification and cloning of the relevant regions of FCRL1 into the vector, pLenti-CMV-GFP-Puro. The current stage of this study is at the transfection and transduction steps, and future steps include B cell activation and analysis by confocal microscopy.

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