Scientists Discover New G-Quadruplex Target for Monkeypox Virus Detection and Therapeutics

Researchers at JNCASR have identified unique DNA structures in MPV genome that enable precise diagnostic and therapeutic strategies.

Scientists from the Jawaharlal Nehru Centre for Advanced Scientific Research (JNCASR), an autonomous institution of the Department of Science and Technology (DST), have identified a new route to understand the virology of Monkeypox Virus (MPV) and develop diagnostic tools for infection detection and potential therapeutic interventions.

The recent outbreak of MPV, now renamed Mpox virus, has been declared a Public Health Emergency of International Concern (PHEIC) twice within the last three years, underscoring the urgent need for rapid and reliable diagnostic and therapeutic solutions. Despite the widespread impact of this outbreak, the transmission mechanisms and clinical symptoms of MPV remain poorly understood, making early detection and effective treatment challenging. To address these issues, scientists are exploring innovative methods to understand the virology of MPV and develop diagnostic approaches that are highly specific and sensitive.

Leveraging G-Quadruplex Structures for Reliable Detection

MPV is a double-stranded DNA (dsDNA) virus, and traditional detection techniques such as Polymerase Chain Reaction (PCR) primarily rely on the amplification of dsDNA using fluorescent probes. However, these methods often struggle to differentiate between specific and non-specific amplification products due to limitations in their sensitivity and ability to identify unique nucleic acid structures. The new discovery focuses on noncanonical DNA structures known as G-quadruplexes (GQs)—unique four-stranded DNA conformations that form in guanine-rich sequences. These GQs, stabilized by hydrogen bonding between guanines, deviate from the classical double-helix structure and offer new potential targets for diagnostic and therapeutic applications.

Scientists at JNCASR have identified and characterized a set of highly conserved GQ-forming DNA sequences within the MPV genome. Utilizing a tailored fluorescent small-molecule probe, the researchers successfully detected a specific GQ sequence in the MPV genome, achieving precise and sensitive identification of the virus. The GQ-forming sequences identified are stable under physiological conditions, highly conserved, and absent in other pox viruses, other pathogens, and the human genome. These features make them ideal targets for developing diagnostic tools and therapeutic strategies.

The newly developed fluorogenic molecular probe, named BBJL, provides a more than 250-fold enhancement in fluorescence output upon binding with MPV GQs. This probe is non-fluorescent in the absence of the target GQ-DNA and represents the first practical demonstration of GQ-targeted diagnostics for MPV detection. BBJL’s ability to selectively identify the highly conserved GQ sequence in the MPV genome sets a precedent for developing detection techniques targeting noncanonical nucleic acids.

Broader Implications for Diagnostics and Therapeutics

The GQ-targeted molecular probe developed by the team, which was originally demonstrated for SARS-CoV-2/COVID-19 detection through the GQ-RCP (G-quadruplex targeted reliable conformation polymorphism) platform, has now been expanded to include MPV detection. This innovative strategy utilizes GQs as molecular targets to overcome challenges in existing amplification-based techniques, which often lead to false-positive results due to non-specific amplification. The probe’s ability to efficiently distinguish MPV-derived GQs from other DNA sequences present in the human genome ensures high specificity and sensitivity.

The identification of these highly conserved GQs within the MPV genome opens up potential avenues for antiviral interventions. The GQ sequences serve as ideal targets for the development of novel therapeutic agents designed to inhibit MPV replication. This could pave the way for a new class of antiviral treatments that specifically target MPV RNA or DNA structures.

The mapping of MPV genome to identify additional GQ targets for future therapeutics is ongoing, which could further expand the range of applications for GQ-based diagnostics and therapeutics. These findings not only enhance our understanding of MPV virology but also offer a pathway to develop more reliable diagnostic tools and antiviral therapies.

Impact on Public Health and Scientific Research

The novel GQ-targeted detection method developed by JNCASR represents a significant breakthrough in MPV diagnostics. By leveraging the unique structural properties of GQs, scientists can overcome existing limitations in traditional PCR-based detection techniques, enabling faster, more accurate, and reliable diagnosis of MPV infections. This innovative approach also holds promise for broader applications in other viral infections, where GQs may serve as potential therapeutic targets.

Furthermore, the integration of GQ-targeted diagnostics with existing MPV surveillance efforts can aid in early detection and containment of outbreaks, thereby reducing transmission risks and protecting public health. The GQ-targeted molecular probe developed by JNCASR sets a new standard for MPV detection and highlights the value of collaborative scientific research in addressing global health challenges.

This groundbreaking work not only advances our understanding of MPV virology but also demonstrates the potential of GQ-targeted diagnostics to revolutionize viral detection and therapeutics. The broader scientific community can now leverage these findings to explore GQ structures as potential antiviral targets, contributing to the development of new, highly effective diagnostic and therapeutic strategies.