Metriotect - Project description

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Endometriosis is a debilitating chronic disease that is anticipated to affect 10-15% of women around the world [1]. This disease affects women of reproductive age and is dependent on estrogen [2,3]. Even in Europe it takes on average 7 years to diagnose it [1]. Women suffering from period pain, bloating, painful sexual intercourse and fatigue are usually undermined while their quality of life and their mental health keeps plummeting [3]. Undiagnosed endometriosis can cause infertility which is the most common reason for diagnosis [1,4].

At the moment, the most valid diagnosis of endometriosis is laparoscopic surgery, which includes making a small incision in the patient's stomach and investigation of any pathologies in the tissues around the pelvic area [5]. In rare cases, when endometriosis is diagnosed early the patient has two options: either to get pregnant (if that is desirable and endometriosis has not affected the ovaries) or start hormonal medication [2]. In later stages of the disease, the patient undergoes surgical removal of the tissue and then is prescribed medication that decreases fertility [2]. The surgery must be conducted by a professional working with reproductive health to avoid leaving the patient infertile [2,5].

Medical professionals are insisting on endometriosis to be considered a public health issue and not an individual problem [3]. Due to increased interest in women's reproductive health and endometriosis, scientists have started investigating molecular mechanisms of endometriosis, that lead our team to investigate potential biomarkers associated with endometriosis [6].


Our team bases our project Metriotect on the newest research in potential miRNA biomarkers in endometriosis patients [7]. To enable these miRNAs, we created a hypothetical workflow. First, we need a patient's blood sample, from which all RNA is isolated, then these RNAs are transcribed and amplified. Our team created an RNA-guide (gRNA) for one of the potential endometriosis biomarkers and engineered dead Cas12f (dCas12f) proteins to enable accurate detection [8]. These dCdas12f proteins not only are able to bind to the target DNA sequence with the help of gRNA but also have antigens attached, one that is able to attach to antibodies on a lateral flow test and one on the other side that acts like a fluorophore – making it visible to the eye to see if the patient has these endometriosis associated biomarkers. We invision this lateral flow test to look like a covid antigen test or a pregnancy test, making it easy, quick and affordable to use.

Our journey

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In 2023 the new iGEM team started assembling. We started of quite strong around 8 people making the team, but as we brainstormed for new project ideas and went through multiple different topics raining from conservation, biomaterials our team decreased significantly. As is now UiOslo Norway iGEM 2023 team is made of 5 people who are studying for their master's degrees in genetics, neuroscience, molecular biology and biochemistry. Metriotect came to be our team's project because all of us agreed of the importance of it for improving women's health and we quickly saw a vision of how to make an early endometriosis diagnosis possible.

Our goals for iGem 2023:

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-Bring awareness of endometriosis and women's reproductive health. Why we think early diagnosis and treatment of patients would improve their lives significantly.

-Create a proof of concept: engineer a construct able to bind to target dsDNA

-While talking with experts in the field working with endometriosis patients (medical doctors), bioethics experts and other scientists working to create a different tool for diagnosis we aimed to integrate and implement their advice and input.

Endometriosis diagnosis now:

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Even though endometriosis occurs quite often, it is estimated that 1 in 10 women suffer from it, the diagnosis can take around 7 years [1]. Patients suffering from chronic pain are easily undermined or they themselves think it is a normal part of their menstrual cycle, early symptoms include pain in the pelvic region, during sexual intercourse following fatigue caused by it [1]. A regular gynecologist appointment usually includes ultrasound checkup which if conducted by a professional with a trained eye can detect early endometriosis stages, this is followed by histological tests and an MRI [1-6]. However, this is usually not enough for a diagnosis and doctors suspecting endometriosis must conduct laparoscopic surgery [1-6]. Laparoscopic surgery requires a professional surgeon that also specializes in reproductive health, it is time and resource consuming in addition to being an invasive diagnostic method [2,6].

What an early diagnosis entails:

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An early diagnosis of endometriosis, first and foremost, would prevent infertility, that is a debilitating result of this disease. Even for women not planning pregnancy it would help prevent pain, fatigue and discomfort in their daily lives – improving their quality of life significantly. Early diagnosis can mean the progression of the disease can be prevented or almost stopped. In early stages of endometriosis, when the disease has not affected the ovaries, the patient can get pregnant or start hormonal contraception [2,4].

Metriotect: our solutions for early endometriosis diagnosis

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Due to current preventative measures we aimed to create a way to diagnose endometriosis early and help women around the world.

By engineering dCas112f proteins with our specific gRNAs we believe that early diagnosis can be made possible.

CRISPR/Cas-based lateral flow assays for nucleic acid detection have emerged during the covid pandemic and inspired us to use a similar approach for endometriosis biomarkers [8,9]. We created two dCas12 proteins that have different purification tags (histidine and streptavidin tags) and can bind gRNA (this way we elongated the detection sequence).

Nucleic acid based diagnosis methods are beneficial because it is possible to amplify them to a necessary input for downstream applications [9]. And even though CRISPR/Cas systems are known for their genetic engineering applications have also been used for sensitive diagnostics methods [9].

Our choice of preamplification for the workflow is RPA – recombinase polymerase amplification, which allows rapid amplification for minimally processed samples and the use of small portable instruments [10]. In this way we cut costs, time and avoid the necessity for highly trained personnel [8,10].

To achieve a more stable biomarker – reverse transcriptase added to the RPA reaction converts RNA to cDNA and the amplified biomarker sequences then are in dsDNA form which is more stable than ssRNA [10]. All this is achieved in a single tube and is way more efficient and cost effective than qPCR based diagnostics [10].