CerviCare
CerviCare, the 2023 project by UMaryland iGEM, harnesses the power of synthetic biology to address the critical issue of limited access to pre cervical cancer screening in underserved regions. Our mission is to develop a revolutionary point-of-care screening tool that leverages upregulated miRNA biomarkers found in the urine of individuals at risk of pre cervical and cervical cancer. This groundbreaking technology produces a color output through the integration of red fluorescent protein, serving as an accessible and visually intuitive indicator. Our ultimate objective is to prompt individuals to seek further medical evaluation, ideally culminating in a confirmatory diagnosis via biopsy.
The CerviCare device aims to prevent unnecessary fatalities of cervical cancer through early intervention and screening. This innovation is especially crucial in areas with limited access to healthcare professionals capable of performing essential procedures like pap smears and biopsies. By democratizing access to cervical cancer screening, we aspire to make a meaningful impact on the health and well-being of individuals in underserved communities.
How it Works
Our project is poised to revolutionize cervical cancer screening by harnessing the combined power of two cutting-edge biosensors: Toehold switches and Synthetic RNA devices. This innovative approach ensures heightened sensitivity and specificity in our screening test. When the targeted miRNAs bind to these sensors, a cascade of conformational changes is triggered, culminating in a clear color output, indicating the presence or absence of the biomarker.
Our vision for the final device includes the incorporation of a sophisticated microfluidic structure. This component will provide distinct color outputs for each of our target miRNAs (miR-21-5p, miR-199a-5p, and miR-155-5p). Through the integration of these state-of-the-art technologies, our goal is twofold: firstly, to offer individuals a prompt indication of the urgency for further medical evaluation, such as a pap smear or biopsy; and secondly, to provide a highly accurate screening test with minimal false negatives or false positives.
By using these advanced biosensors and microfluidic systems, we aim to enhance the effectiveness and accessibility of cervical cancer screening, ultimately contributing to early detection and improved outcomes for individuals at risk, particularly those in underserved regions.
Construct Design
1. Toehold Report
2. Ribozyme Report
3. miRNA gBlocks Design(s)
We designed gBlocks encoding for our distinct miRNA biomarkers (miR-21-5p, miR-199a-5p, and miR-155-5p). The overarching goal of this endeavor was to develop a specialized miRNA plasmid capable of producing our unique miRNA biomarkers. These biomarkers would serve as targets for our toehold switch system, ultimately triggering the production of an mRFP signal.
To streamline our efforts, we chose to work with the plasmid pSB1A3-Amp-mRFP (Fig 1.), which was readily available in our laboratory resources. This decision was primarily motivated by practical considerations and resource efficiency.
The miRNA plasmid gBlocks were designed to include recognition sequences for the EcoRI and SpeI restriction enzymes (Fig 2.). Our rationale behind this design choice was to facilitate the precise excision of the mRFP sequence from our plasmid. This step was crucial because our toehold switch designs already incorporated the mRFP component. It is essential to ensure that our toehold switch system is the only system producing a signal in order to observe whether or not the system is functioning when our biomarkers are present.
References
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Following the design of the gBlocks, we conducted the vital step of creating restriction maps (Fig 3.). These maps played a pivotal role in verifying the correct alignment and presence of the EcoRI and SpeI restriction enzyme recognition sites within our sequence. This meticulous planning and validation process were undertaken to ensure the accuracy and success of our research.