Cells produce different versions of genes from the same genetic material. This ability is crucial for the functioning of organisms in various situations. However, current methods for studying different gene versions are limited and cannot precisely analyze their expression within the organism. Additionally, studying different protein versions is challenging because existing tools may not distinguish between them. While there are techniques available to measure the abundance of gene versions, they require isolating and separating cells, which results in the loss of important information about cell interactions and the environment. In this project, the investigators aim to develop a new technology called electromicrofluidics (EMF), which combines electricity, engineered gels, and cell culture on a microchip to automatically study gene versions at the single-cell level. This technology will provide insights into how these different gene versions function in complex biological processes involving multiple cells, such as development, growth, and repair. In addition to creating a novel biosensing approach, the research will be integrated across several educational fronts, including graduate program development, active learning through student-designed projects, social media dissemination, undergraduate and minority research opportunities,and outreach programs.The research aims to establish an EMF based biosensing platform for comprehensive gene expression profiling. This platform incorporates electroalignment-enhanced anisotropic gel polymerization and in situ immunolabeling and amplification to enable simultaneous analysis of multiple gene expression modes. The investigators will explore different gel layer configurations suitable for studying protein isoforms and (ribonucleic acid) RNA splicing variants. This approach allows for accurate profiling while minimizing sample loss and contamination during transfer. By implementing electroalignment-enhanced anisotropic gel polymerization, the lateral diffusion of molecules is constrained, resulting in improved spatial resolution and sensitivity. This enhancement facilitates in situ single-cell analysis, enabling the study of spatial coordination and cell heterogeneity. Furthermore, the use of EMF technology automates the assay procedures, enhancing reproducibility and facilitating wider adoption of the platform. Initially, 3D gel assays will be developed to analyze protein isoforms and RNA splice variants. Subsequently, these assays will be integrated into a versatile platform capable of simultaneously detecting multiple protein isoforms and RNA splicing variants. The project will focus on mapping the Neurogenic locus notch homolog protein (NOTCH) family members in migrating cell monolayers to evaluate their regulatory functions in collective cell migration.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
|Effective start/end date
|7/1/23 → 6/30/26
- National Science Foundation: $300,000.00
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Chembe hutokeza matoleo tofauti-tofauti ya chembe za urithi kutoka kwa chembe za urithi zilezile. Uwezo huo ni muhimu sana kwa utendaji wa viumbe katika hali mbalimbali. Hata hivyo, mbinu za sasa za kuchunguza matoleo tofauti ya jeni ni ndogo na haziwezi kuchanganua kwa usahihi usemi wao ndani ya kiumbe. Kwa kuongezea, kuchunguza matoleo tofauti ya protini ni changamoto kwa sababu zana zilizopo zinaweza kutofautisha kati yao. Ingawa kuna mbinu zinazopatikana kupima wingi wa matoleo ya jeni, zinahitaji kutenganisha na kutenganisha seli, ambayo husababisha upotezaji wa habari muhimu kuhusu mwingiliano wa seli na mazingira. Katika mradi huu, watafiti wana lengo la kuendeleza teknolojia mpya inayoitwa electromicrofluidics (EMF), ambayo inachanganya umeme, jeli za uhandisi, na utamaduni wa seli kwenye kipande cha microchip, kwa kujifunza moja kwa moja matoleo ya jeni katika kiwango cha seli moja. Teknolojia hii itatoa ufahamu juu ya jinsi matoleo haya tofauti ya jeni yanavyofanya kazi katika michakato tata ya kibiolojia inayohusisha seli nyingi, kama vile maendeleo, ukuaji, na ukarabati. "Kwa kuongezea kuunda mbinu mpya ya biosensing, utafiti utaunganishwa katika pande kadhaa za elimu, pamoja na maendeleo ya programu ya kuhitimu, kujifunza kwa bidii kupitia miradi iliyoundwa na wanafunzi, kueneza media ya kijamii, fursa za utafiti wa shahada ya kwanza na wachache, na programu za ufikiaji. "" - Biosensing." Programu hii inajumuisha electroalignment-kuimarishwa anisotropic gel polymerization na in situ immunolabeling na amplification kuwezesha uchambuzi wa wakati mmoja wa modes nyingi za usemi wa jeni. Kwa hivyo, watafiti watachunguza muundo tofauti wa tabaka la gel inayofaa kwa kusoma isoforms za protini na (asidi ya ribonucleic) tofauti za splicing ya RNA. Njia hii inaruhusu kwa profiling sahihi wakati kupunguza sampuli hasara na uchafuzi wakati wa uhamisho. Kwa kutekeleza electroalignment-kuimarishwa anisotropic gel polymerization, lateral kuenea ya molekuli ni vikwazo, na kusababisha kuboreshwa nafasi azimio na unyeti. Kuboresha hii kuwezesha in situ single-seli uchambuzi, kuwezesha utafiti wa uratibu wa nafasi na seli heterogeneity. Kwa kuongezea, matumizi ya teknolojia ya EMF hufanya utaratibu wa kupima, kuimarisha reproducibility na kuwezesha kupitishwa kwa jukwaa. Kwa mfano, katika kesi ya 3D, vipimo vya gel vinaweza kutumiwa kwa ajili ya kuchunguza aina za protini na aina za splicing za RNA. Baadaye, vipimo hivi vitaunganishwa katika jukwaa la kubadilika linaloweza kugundua wakati huo huo isoforms nyingi za protini na tofauti za splicing za RNA. "Mfano wa ""neurogenic locus notch homolog protein"" (NOTCH) ni sehemu ya familia ya ""neurogenic locus notch homolog protein"" (NOTCH) ambayo inahusika katika kuharakisha na kuharakisha shughuli za kibiolojia za chembe za urithi, na inahusu uwezo wa kuharakisha shughuli za kibiolojia za chembe za urithi." "Mwaka wa ""Mwaka wa Utafiti"" wa 2017 - 2017: ""Mwaka wa Utafiti"" wa 2017 - 2017: ""Mwaka wa Utafiti"" wa 2017 - 2017: ""Mwaka wa Utafiti"" wa 2017 - 2017: ""Mwaka wa Utafiti"" wa 2017 - 2017" Labels hizi ni yanayotokana kulingana na tuzo msingi / misaada. Zikiwa pamoja hufanyiza alama ya kidole ya pekee.