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In Situ Sequencing of Molecular Interactomes

日期: 2015-06-02

生命学院学术报告

题目:In Situ Sequencing of Molecular Interactomes

报告人:Liangcai Gu, Ph.D.

Research Fellow of Genetics, Harvard Medical School

Assistant Professor of Biochemistry, University of Washington

时 间:2015年6月5日,10:30 am-11:30 am

地 点:william hill中文网411会议室

联系人:昌增益 (changzy@pku.edu.cn)

Abstract:

We developed a single-molecular-interaction sequencing (SMI-seq) technology to improve quality and cost-effectiveness of large-scale molecular interaction analyses—in particular, protein–protein interaction profiling. SMI-seq involves attaching proteins to a DNA identifier or `barcode` via ribosome display or enzymatic conjugation, assaying barcoded proteins en masse in aqueous solution and immobilizing them in a polyacrylamide thin film. Barcoding DNAs are then in situ amplified and sequenced to quantitate protein-protein interactions. SMI-seq enables precise protein quantification and one-pot ‘library vs. library’ screening to simultaneously interrogate molecular binding affinity and specificity. We demonstrated its applications in G-protein coupled receptor drug screening and antibody–antigen binding profiling. I will discuss the possibility of expanding in situ sequencing to analyze non-barcoded proteins in cells, for example, the profiling of vastly diverse antigen receptor–ligand interactions and cell responses upon receptor activation that are important to adaptive immunity.

Reference

Gu et al., (2014). Multiplex single-molecule interaction profiling of DNA-barcoded proteins. Nature, 515:554-557.

欢迎各位老师同学积极参加!