INTEGRATION OF RNAI INTO THE C. ELEGANS GENOME TO IMPACT DAT-1EXPRESSION

Abstract:

Addiction is an epidemic that has overtaken the United States of America since the 1990s and has seen a rise in deaths since, with synthetic drugs such as Fentanyl flooding the market. It overtakes those we know all around us, whether it is apparent or not. In the United States alone, there were over 40 million registered cases of SUDs (substance use disorders) in 2020, and only 6.5% of them received treatment of any kind (National Institute on Drug Abuse, 2024). The effects of addiction have cost the United States nearly 700 billion dollars annually (Addiction help, 2024), and almost 100,000 people die per year. There have been many different treatment measures, such as Naloxone, Methadone, Buprenorphine, and nicotine replacement therapies, but there has not been much curing of genetic mutations or one of the roots of addiction: dopamine.

Recently, in Brazil, there has been a breakthrough in vaccines using a non-protein-based, V4N2 molecular-based treatment for crack cocaine addiction, which is a promising step in the treatment of addiction (National Library of Medicine, 2023). We have traced the dopamine gene, SLC6A3 (DAT-1), as a reuptake from the synapse that is highly linked to addiction (Neuropsychopharmacology, 2007) to substances such as cocaine and alcohol, to name a few. Time and time again, dopamine receptors have been linked to addiction, but we wanted to study the effects of dopamine levels on a subject. Theoretically, if you can lower levels of dopamine in humans, there shouldn’t be that “want” to reach for drugs that addiction has.

However, many view it to be a crime-ridden disease that only affects the poor and marginalized. Addiction’s stigmatized viewpoint has assumed addicts to be not a part of society, with the government recently withdrawing 9 billion dollars from the National Institute of Health, with targets such as addiction, gun violence, and PTSD (Health Policy Watch, 2025).

Opposed to that, methods such as CRISPR Cas9, Zinc Finger, or RNA Interference have increased in popularity, and gene editing has made a scientific breakthrough, including a cure for Sickle Cell Anemia (Columbia University Irving Medical Center, 2021). Dopamine levels in addicts are commonly higher than in the average person, around 10 times higher when on substances (Yale Medicine, 2024). Among most users, the rush they feel from using floods the brain and nervous system with dopamine and serotonin.  But what if we silenced genes that cause problems and, one day, silenced mutations?

In our study, we look toward using a strain of C. elegans OK157 that is modified to contain the DAT-1 gene, consuming transformed E. coli HT115 with the L4440 Plasmid to silence the DAT1 gene prevalent in them. C. elegans is an excellent model organism to use due to its straightforward anatomy and similarity to humans. “Emerging evidence suggests that C. elegans is an excellent model to identify molecular mechanisms that mediate drug-induced behavior and potential targets for medication development for various addictive compounds” (Science Direct, 2016).

With our reflection on addiction in the procedure of C. elegans, we aim to silence the DAT-1 gene to show if it is possible to lower dopamine in humans one day.

Full Paper: https://docs.google.com/file/d/1_NUsYyy3B3dFR140a44-CfCojYX-muHr/edit?usp=docslist_api&filetype=msword

Authors: Jake McClure, Jorja Lamley, and Alexis Kendrick