Ekbal (Eke) Djibril
Eke, a freshman studying Health Science at Howard University, optimized RNA stability for its use in a new and innovative infectious disease diagnostic tool called CARMEN.
Current diagnostic tools need to meet a wide range of public health needs. CARMEN, a diagnostic platform that delivers high throughput comprehensive information, can test up to 192 patient samples for up to 24 different pathogens, including Ebola, Yellow Fever Virus, HIV, and more simultaneously using a microfluidic system, delivering results in less than a day. I came into the Broad with a rigid plan for my career goals in a manner that would not allow deviation from these plans or make room for anything not on there. At the end of this program, I learned a valuable lesson that you should let things come to you and not always seek things out, I can happily say I got rid of this strict and rigid plan because life is going to life, and you can’t always control every step of it. BSRP, being my first undergraduate experience as a freshman, was the biggest blessing I could have received because I left with so much more than an understanding of research but with a family. A family of mentors, friends, and people who want to see me succeed in a society that tells me I am not meant for these spaces. This program gave me the opportunity to understand how research questions are developed, how to communicate scientific concepts, and how to be more independent in the lab. For that, I am forever grateful for this life-changing opportunity, and I am excited to see where my journey takes me as a developing scientist who welcomes deviations!
However, RNA is inherently unstable; an unstable RNA input could lead to inaccurate and unreliable results. We have tried to combat this by introducing transfer RNA into our RNA samples to act as a buffer during storage. However, we must also ensure that the added tRNA does not interfere with the highly sensitive Cas enzyme trans cleavage mechanism used in CARMEN. Experimentally, Yellow Fever Virus RNA was suspended in water with or without tRNA and placed in five different temperature storage conditions (-80ºC, -20ºC, 4ºC, 25ºC, and 37ºC). At each sampling time point (ranging from Day 0 to Week 4), an aliquot of RNA from each storage condition was collected and serially diluted to produce a standard curve on qPCR, allowing comparison of RNA degradation over time. Our results indicate that tRNA has a protective quality and helps stabilize the viral RNA at temperatures as high as 37ºC. Using the 37ºC data as an accelerated stability condition, we can predict the stability of RNA with tRNA stored at various temperatures. Secondly, adding tRNA does not affect the Cas trans cleavage mechanism, maintaining the sensitivity of CARMEN’s blood-borne pathogen assay. Ultimately, we aim to share this diagnostic tool with our partners in West Africa through project SENTINEL, which allows them to save money and time where resources are already limited.
Project: Optimizing RNA Stability for Diagnostics in West African Countries
Mentors: Al Ozonoff, Elyse Stachler & Kyle Pacheco
PI: Sabeti Lab