{"693760":{"#nid":"693760","#data":{"type":"event","title":"BioE PhD Proposal Presentation-  Shifeng Xu ","body":[{"value":"\u003Cp\u003E\u003Cstrong\u003EAdvisor:\u003C\/strong\u003E\u0026nbsp;Dr. Mark Styczynski (School of Chemical and Biomolecular Engineering)\u003C\/p\u003E\u003Cp\u003E\u003Cstrong\u003ECommittee Members:\u003C\/strong\u003E\u003C\/p\u003E\u003Cp\u003EDr. John Blazeck (School of Chemical and Biomolecular Engineering)\u003C\/p\u003E\u003Cp\u003EDr. Julie Champion (School of Chemical and Biomolecular Engineering)\u003C\/p\u003E\u003Cp\u003EDr. Brian Hammer (School of Biological Sciences)\u003C\/p\u003E\u003Cp\u003EDr. Valeria Milam (School of Materials Science and Engineering)\u003C\/p\u003E\u003Cp\u003E\u0026nbsp;\u003C\/p\u003E\u003Cp\u003E\u003Cstrong\u003EEngineering Modular Target Recognition for Cell-Free Biosensing in One-Pot PURE\u003C\/strong\u003E\u003C\/p\u003E\u003Cp\u003EPoint-of-care (POC) diagnostics benefit from low-cost biosensing platforms that offer simple readouts and can be readily adapted to detect diverse analytes. Cell-free protein synthesis (CFPS) offers a programmable environment for combining molecular recognition elements and genetic circuits for biosensing. One such approach, the T7 RNA polymerase-Linked ImmunoSensing Assay (TLISA), uses nanobodies to reassemble split polymerase fragments upon simultaneous binding to a target protein, driving reporter expression. A more sensitive version of TLISA has been implemented in PURE, a CFPS system reconstituted from purified components. However, commercial PURE costs $1,110 per mL of reaction, making biosensor optimization expensive and limits affordability for POC deployment. One-pot PURE simplifies preparation by jointly purifying the 36 PURE proteins from a co-culture, reducing costs to $90 per mL. I hypothesize that one-pot PURE will improve TLISA\u2019s affordability, while interchangeable nucleic acid recognition elements will enable modular detection of protein and nucleic acid targets. These elements would address a separate limitation: adapting TLISA to new targets requires cloning new affinity domain\u2013polymerase fusions, slowing sensor development. First, I will establish and optimize one-pot PURE in-house and validate its support for TLISA sensing. Second, I will engineer an RNA hairpin-aptamer TLISA architecture that tethers target-binding aptamers to split polymerase fragments through specific protein-RNA interactions. This design would allow target specificity to be changed by switching out aptamers while retaining the same polymerase fusion proteins, reducing the need for cloning. Third, I will adapt TLISA to detect microRNA, extending the platform to sense nucleic acid biomarkers. Together, this work aims to establish an affordable, modular cell-free biosensing platform that accelerates sensor development and supports future POC diagnostic applications.\u003C\/p\u003E","summary":"","format":"limited_html"}],"field_subtitle":"","field_summary":[{"value":"\u003Cp\u003EBioE PhD Proposal Presentation- \u0026nbsp;\u0022Engineering Modular Target Recognition for Cell-Free Biosensing in One-Pot PURE\u0022 - Shifeng Xu\u0026nbsp;\u003C\/p\u003E","format":"limited_html"}],"field_summary_sentence":[{"value":"\u0022Engineering Modular Target Recognition for Cell-Free Biosensing in One-Pot PURE\u0022"}],"uid":"27917","created_gmt":"2026-11-04 15:39:32","changed_gmt":"2026-11-04 15:41:06","author":"Laura Paige","boilerplate_text":"","field_publication":"","field_article_url":"","field_event_time":{"event_time_start":"2026-11-16T13:30:00-05:00","event_time_end":"2026-11-16T15:30:00-05:00","event_time_end_last":"2026-11-16T15:30:00-05:00","gmt_time_start":"2026-11-16 18:30:00","gmt_time_end":"2026-11-16 20:30:00","gmt_time_end_last":"2026-11-16 20:30:00","rrule":null,"timezone":"America\/New_York"},"location":"5029 EBB","extras":[],"groups":[{"id":"65448","name":"Bioengineering Graduate Program"}],"categories":[],"keywords":[{"id":"172056","name":"go-BioE"}],"core_research_areas":[],"news_room_topics":[],"event_categories":[],"invited_audience":[],"affiliations":[],"classification":[],"areas_of_expertise":[],"news_and_recent_appearances":[],"phone":[],"contact":[],"email":[],"slides":[],"orientation":[],"userdata":""}}}