Current detection method for colorectal cancer includes colonoscopy and fecal immunochemical test (FIT), which both have some limitations.

    Limitation of colonoscopy

1. Invasive Examination

Colonoscopy is an invasive procedure that involves inserting a flexible tube with a camera (colonoscope) into the rectum and colon. This can cause discomfort, cramping, and even pain for some patients, especially during the insertion and navigation of the scope. The invasive examination led to a low willingness for public to participate, resulting in patients delaying seeking medical care.

2. Potential Risk of Acute Physical Hazard

Nonetheless, colonoscopy carries some risks, including bleeding, bowel perforation, or infection. Perforation can occur when the colon's lining is punctured, potentially leading to infection and requiring surgical repair. These complications can be severe and may require immediate medical attention.

3. High Financial Burden

Financial burden is another issue; colonoscopy can be relatively expensive, especially for patients without insurance coverage. The cost includes not only the procedure itself but also the facility, anesthesia, and pathology fees for analyzing the collected tissue samples. According to the Center for Health and Cancer Prevention Institute, the total cost of a colonoscopy is $850 USD.

4. Dietary Restriction

A low-fiber diet is required on the day prior to the colonoscopy examination. Reducing the uptake of fiber can effectively reduce bowel movements, which makes the enema process more comprehensive. A low-fiber diet should avoid consuming soybeans, whole grain products, raw vegetables, red meat, etc., which causes inconvenience and could lower examination willingness.

    Limitation of Fecal Immunochemical Test

The fetal immunochemical test (FIT), while non-invasive, may produce false-negative results, especially in cases where bleeding is intermittent or at a low level. This can lead to delayed diagnosis and missed opportunities for early intervention. According to the reference paper (https://aacrjournals.org/clincancerres/article/23/8/2061/123555/Fecal-Bacteria-Act-as-Novel-Biomarkers-for), the FIT test has low sensitivity at the first stage of colorectal cancer, which may delay examination. The FIT test can also be influenced by dietary choices and certain medications. Some foods, such as red meat, raw vegetables, and vitamin C supplements, can lead to false-positive or false-negative results due to their impact on gastrointestinal bleeding.

    Our Novel Detection Method: NanocircDx

Recognizing the limitations and drawbacks associated with traditional cancer detection methods like colonoscopy and the Fecal Immunochemical Test (FIT), we are excited to introduce a groundbreaking approach called NanocircDX.

1. Enhanced Speed Detection in Cancer Early Stage

Our primary aim with NanocircDX is to revolutionize cancer detection by offering a faster and more accurate method, particularly for detecting cancer at its earliest and most treatable stages.

2. Minimal-Invasive Testing

Unlike invasive procedures such as colonoscopy, NanocircDX offers a minimal-invasive testing process. Patients will no longer need to endure uncomfortable or embarrassing procedures, increasing their willingness to participate in regular screenings.

3. Improved Patient Experience

The simplicity and minimal-invasiveness of NanocircDX lead to a more positive and comfortable patient experience. This encourages individuals to proactively engage in cancer screening, promoting early detection and better outcomes.

4. Quick Results

NanocircDX offers rapid results within 2.5 hours, reducing the waiting time for patients. Early detection can lead to early intervention, potentially saving lives and reducing the overall burden of cancer treatment.

5. Enhance Public Willingness

NanocircDX is designed not only to be a more effective detection method but also to encourage more individuals to prioritize their health through regular screenings. By eliminating the discomfort and anxiety associated with traditional screening methods, NanocircDX aims to make cancer screening a less intimidating experience.

Recognizing the scarcity of comprehensive circRNA synthesis protocols in the current literature, we have successfully developed a complete detection process. This valuable resource is intended to assist future iGEM teams and researchers interested in circRNA detection. Our well-defined synthesis steps ensure precise and efficient circRNA production, addressing the challenges associated with the circRNA detection process.

In Vitro Transcription

Prior to the circularization process, users should prepare the target sequence. The selected sequence types could either be DNA or ssRNA. If the prepared sequence is in DNA form, then the sequence should require the T7 promoter and cDNA of circRNA, and then conduct IVT (in vitro transcription) to transcribe the sequence into ssRNA. If the prepared sequence is in ssRNA form, then users should skip IVT and DNase I and do the following process:

DNase I

In this step, DNase I is added to degrade template DNA for the IVT process.

RNA Clean-up

RNA clean-up is required to remove enzymes, salts, and contaminants. This purification process enhances the quality and integrity of RNA, ensuring accurate downstream applications.

Monophosphorylation

Monophosphorylation is aimed to substitute triphosphate with single phosphate group on a sequence. Pyrophosphatase is the common used enzyme in monophosphorylation process. In RNA circularization, pyrophosphatase is typically applied to 5’ end of the RNA molecule, enabling the formation of a circular structure by covalently linking the monophosphorylated end to the other end.

RNA Clean-up

RNA clean-up is required to remove enzymes, salts, and contaminants. This purification process enhances the quality and integrity of RNA, ensuring accurate downstream applications.

Form Nicked dsRNA

In this step, splint and ssRNA are introduced, and after a high-temperature reaction, a slow cooling process promotes annealing, resulting in the formation of dsRNA. The formed dsRNA assists in the subsequent circularization step.

Circularization

T4 RNA ligase 2 is an enzyme commonly used in molecular biology for various RNA manipulation techniques, including circularization. When applied to circularization, T4 RNA ligase 2 catalyzes the formation of a covalent bond between the 3' and 5' ends of an RNA molecule, resulting in a circular RNA structure.

DNase I

In this step, DNase I is added to degrade template DNA for the IVT process.

RNA Clean-up

RNA clean-up is required to remove enzymes, salts, and contaminants. This purification process enhances the quality and integrity of RNA, ensuring accurate downstream applications.

RNase R

RNase R is a kind of exoribonuclease enzyme used to degrade the exposed end of RNA. It has specificity for linear-form RNA and has circRNA resistance. By adding RNase R to the circularization product, unsuccessful circularization RNA could be degraded, leaving a pure circRNA product.

RNA Clean-up

RNA clean-up is required to remove enzymes, salts, and contaminants. This purification process enhances the quality and integrity of RNA, ensuring accurate downstream applications.

The shortcomings of many current detection methods, which rely on complicated manual operation and interpretation, have led to concerns about increased human errors and uncertainties in sample analysis. These limitations can potentially impact the accuracy of the interpretation and the overall reliability of the results. To address these challenges, we have developed an innovative detection device that streamlines the entire process, reducing the need for manual intervention and offering a user-friendly experience.

Unlike manual methods that require constant human intervention at various stages, our device is designed to automate the entire detection process. Once a sample is placed into the device, it automatically initiates pre-programmed protocols for different types of sample analyses. Users do not need to provide instructions for each step; the device follows a predetermined sequence based on the type of analysis required. This simplifies the operation and ensures that the process is carried out correctly every time.

To enhance user monitoring and interaction, our device features a user-friendly LED display panel. This panel provides real-time information about the current steps in the analysis process. Users can easily track the progress, ensuring that they are informed and in control throughout the procedure.

We aim to enhance the accuracy, reliability, and user experience of sample detection across various applications, from medical diagnostics to research endeavors. This device marks a crucial step toward more streamlined and error-resistant sample analysis methods. It is intuitive and easy to use, making it accessible to a wide range of users, including healthcare professionals and researchers.

Creating a database for PCRD and gold nanoparticle detection results is a valuable initiative for streamlining the analysis and interpretation of data from different samples. This database allows for faster and more accurate automated analysis of newly imported data, contributing to improved precision in result interpretation.

The database serves as a centralized repository for storing PCRD and gold nanoparticle detection results from various samples. This centralization facilitates efficient data management and retrieval. Users can rapidly access historical data and compare it with newly imported information. This quick search capability streamlines the analysis process, saving time and effort.

Establishing a database for PCRD and gold nanoparticle detection results is a strategic investment in data management and analysis. It enhances the efficiency and accuracy of result interpretation, aids in quality control, and supports research efforts. By leveraging historical data and integrating machine learning, organizations can achieve faster and more reliable insights from their PCRD and gold nanoparticle detection experiments, ultimately advancing their scientific and analytical capabilities.

We designed an APP for accurate identification of post-testing products. Its core functionality is based on a database of testing products, utilizing machine learning technology for identification and analysis.

Firstly, users can capture or upload images of PCRD or gold nanoparticle test results through the application. Subsequently, the application undergoes image processing to extract crucial features and data. These features and data are then transmitted to the testing product database for comparison and identification. The testing product database comprises an extensive collection of reference data from PCR tests and gold nanoparticle tests, including known positive and negative results as well as variations in various contexts. Machine learning models use this reference data for training to accurately identify new results.

Ultimately, users will see the outcome of the application displaying the results of the testing products. Users can trust the results provided by this tool without requiring in-depth expertise.

Furthermore, in the future, this application may include additional features such as saving historical results, providing recommendations or suggesting next steps, and the option to share results with healthcare professionals. This will assist users in better managing their test outcomes and taking the necessary actions promptly. This application is poised to have a significant impact on scientific research, clinical testing, and personal health management.

To help foster cohesion within the iGEM community, NTHU_Taiwan has created a 3D interactive iGEM map. This map allows users to easily click on icons representing iGEM teams and access information about them, contributing to the following:

1. Quick Access to Information

By clicking on icons, users can swiftly access information about other iGEM teams, including their projects, achievements, contact details, and more. This reduces the time spent searching for information, making it easier for members to get to know each other.

2. Establishing Connections

The map can include contact information for teams, facilitating the establishment of connections and communication among members. Such communication aids in building long-term relationships, promoting collaboration, and sharing knowledge.

3. Promoting Community Engagement

Such a map can also highlight events, conferences, and gatherings within the iGEM community, encouraging members to participate in these activities and strengthening community cohesion.

Building cohesion within the iGEM community is a crucial aspect of driving research and development in synthetic biology. By providing easy access to information and promoting collaboration among members, we can create a more robust and collaborative iGEM community, thereby advancing innovation and development in the field of synthetic biology