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Full-Text Articles in Chemistry
A General Approach For Receptor And Antibody-Targeted Detection Of Native Proteins Utilizing Split-Luciferase Reassembly, Cliff I. Stains, Jennifer L. Furman, Jason R. Porter, Srivats Rajagopal, Richard T. Wyatt, Indraneel Ghosh
A General Approach For Receptor And Antibody-Targeted Detection Of Native Proteins Utilizing Split-Luciferase Reassembly, Cliff I. Stains, Jennifer L. Furman, Jason R. Porter, Srivats Rajagopal, Richard T. Wyatt, Indraneel Ghosh
Cliff Stains Publications
The direct detection of native proteins in heterogeneous solutions remains a challenging problem. Standard methodologies rely on a separation step to circumvent nonspecific signal generation. We hypothesized that a simple and general method for the detection of native proteins in solution could be achieved through ternary complexation, where the conditional signal generation afforded by split-protein reporters could be married to the specificity afforded by either native receptors or specific antibodies. Toward this goal, we describe a solution phase split-luciferase assay for native protein detection, where we fused fragmented halves of firefly luciferase to separate receptor fragments or single-chain antibodies, allowing …
Toward A General Approach For Rna-Templated Hierarchical Assembly Of Split-Proteins, Jennifer L. Furman, Ahmed H. Badran, Oluyomi Ajulo, Jason R. Porter, Cliff I. Stains, David J. Segal, Indraneel Ghosh
Toward A General Approach For Rna-Templated Hierarchical Assembly Of Split-Proteins, Jennifer L. Furman, Ahmed H. Badran, Oluyomi Ajulo, Jason R. Porter, Cliff I. Stains, David J. Segal, Indraneel Ghosh
Cliff Stains Publications
The ability to conditionally turn on a signal or induce a function in the presence of a user-defined RNA target has potential applications in medicine and synthetic biology. Although sequence-specific pumilio repeat proteins can target a limited set of ss- RNA sequences, there are no general methods for targeting ssRNA with designed proteins. As a first step toward RNA recognition, we utilized the RNA binding domain of argonaute, implicated in RNA interference, for specifically targeting generic 2-nucleotide, 3′ overhangs of any dsRNA. We tested the reassembly of a split-luciferase enzyme guided by argonaute-mediated recognition of newly generated nucleotide overhangs when …