The Era of Easy Creation of Eco-Friendly Pesticides: Algorithm of ‘Genetic Zipper’ Method in Action

1. Introduction

The continued use of conventional chemical pesticides in agriculture has contributed significantly to crop productivity over the past century. However, this heavy reliance on broad-spectrum chemicals has led to serious environmental and ecological consequences, including the contamination of soil and water, the decline of beneficial insect populations, and the development of resistance in many target pest species. In recent years, global regulations and consumer demand have placed growing pressure on the agricultural sector to find more sustainable and eco-friendly alternatives to synthetic pesticides. These alternatives must ideally combine high efficacy with selectivity, safety for non-target organisms, and environmental biodegradability. Biological control agents and plant-derived compounds are among the traditional alternatives, but they often face limitations in terms of field stability, slow action, or inconsistent performance (Villavicencio-Vásquez et al., 2025; Karuppiah et al., 2025). The next generation of pest control strategies is now turning toward molecular and genetic approaches, particularly those involving nucleic acids. These biotechnological methods allow for species-specific interference with essential physiological processes in pests, offering the potential to revolutionize plant protection while reducing collateral damage to the environment. Importantly, such innovations aim not only to suppress pests effectively but also to do so in a way that aligns with integrated pest management (IPM) frameworks and global sustainability goals. Therefore, the search for innovative and efficient nucleic acid-based pesticides has emerged as a key focus of agricultural research (Kumar et al., 2025; Oberemok et al., 2025a; Oberemok et al., 2025b).

Among the most promising advances in nucleic acid-based pest control are antisense technologies, which act by targeting mature rRNA and/or pre-RNA to disrupt expression in a highly specific manner. Technologies like RNA interference (RNAi), CRISPR/Cas gene editing, and more recently, the Contact Unmodified Antisense DNA biotechnology (CUADb), also known as the ‘genetic zipper’ method, are being actively explored for their pest suppression potential (Kumar et al., 2025). Because of very efficient and easy algorithm, DNA-guided ‘genetic zipper’ method (CUAD biotechnology) is a unique and very potent alternative to other approaches in plant protection based on duplexes of unmodified nucleic acids and RNA-guided nucleases: RNA interference and CRISPR/Cas9 (Figure 1).

Each of these technologies employs sequence-specific recognition, where a guide nucleic acid molecule binds to a complementary rRNA sequence in the pest, leading to insect death. While RNAi and CRISPR/Cas offer powerful gene manipulation capabilities, they often require complex delivery systems, modified molecules, or additional protein components. In contrast, CUADb stands out for its simplicity and low production costs, as it utilizes short, unmodified single-stranded DNA oligonucleotides that can be synthesized rapidly without the need for transgenic components. CUADb works by forming a DNA-rRNA duplex that halts ribosome function and triggers degradation of the target rRNA via DNA-guided rRNase, such as RNase H1 (Oberemok et al., 2025a), ultimately leading to lethal disruption of protein synthesis in the pest (Gal’chinsky et al., 2024; Oberemok et al., 2025a). This contact-based mode of action allows CUADb to be applied as a foliar spray, directly on pests, functioning effectively without needing to be taken up systemically by the plant or pest. Moreover, its rapid design, high selectivity, and minimal risk to non-target organisms make it a compelling alternative to traditional insecticides. As such, CUADb represents a unique intersection of molecular precision and practical agricultural utility.

The ‘genetic zipper’ method has shown notable success in laboratory trials, particularly against hemipteran pests such as aphids, whiteflies, and psyllids, as well as spider mites (Oberemok et al., 2024a; Gavrilova et al., 2025). Experimental evidence suggests that CUADb-based oligonucleotide pesticides can cause 80 to 90 percent mortality in larval or juvenile stages of pests within two weeks of contact exposure (Oberemok et al., 2024a; Gavrilova et al., 2025). One of the most attractive features of this method is its flexibility: using freely accessible digital tools like the DNAInsector platform, users can generate oligonucleotide pesticide sequences tailored to the rRNA of nearly any insect species within seconds (DNAInsector, 2025). This capability allows for rapid and personalized pesticide design, moving pest management away from a one-size-fits-all model toward a more responsive, algorithm-driven approach. The method has also been shown to work across a wide range of economically important species, including those with diverse host plant associations and geographic distributions. Because it relies on conserved rRNA regions (Smit et al., 2007; Noller et al., 2022), it can be applied to multiple related pest species with high confidence in its efficacy. Furthermore, CUADb pesticides are biodegradable and leave no long-term chemical residues, making them ideal for use in organic and environmentally sensitive farming systems (Gal’chinsky et al., 2023; Oberemok et al., 2024b). These combined advantages place CUADb at the forefront of next-generation plant protection strategies. However, as with any emerging technology, careful evaluation of its scalability, regulatory acceptance, and delivery mechanisms in field conditions is essential for its widespread adoption.

This opinion article aims to provide a comprehensive overview of the CUADb-based ‘genetic zipper’ method and its applications in pest control. We begin by explaining the mechanistic basis of this technology and its comparison with other nucleic acid-based approaches, followed by practical aspects of oligonucleotide pesticide design and application. To demonstrate its ease of use and broad applicability, we present 12 DNA sequences with high pesticidal potential designed for insect pests and one mite species representing five continents. We also assess the unique benefits of this method in terms of safety, cost-effectiveness, and species specificity. Finally, we discuss the current limitations and challenges facing CUADb and explore its future potential within integrated pest management systems. By highlighting its scientific foundation and practical promise, this review contributes to the growing discourse on sustainable, algorithm-assisted pest management solutions.

2. Mechanism of the ‘Genetic Zipper’ Method

2.1. CUAD Biotechnology: Step-by Step Action

• Step 1: Arrest of rRNA function, leading to hypercompensation.
• Step 2: Enzymatic degradation of rRNA by DNA-guided rRNase, such as rRNase H1 (Gal’chinsky et al., 2024; Kumar et al., 2025).

DNA containment mechanism (DNAc), a two-step process identified in sternorrhynchans. This process occurs primarily in the nucleus—specifically, the nucleolus—where ribosome biogenesis takes place. In the first step, the target rRNA is functionally ‘arrested,’ leading to its hypercompensation. In the second step, the target rRNA undergoes degradation mediated by DNA-guided rRNases, such as RNase H1. Formation of a duplex between the olinscide and rRNA mimics a zipper mechanism, effectively shutting down rRNA expression and resulting in pest death (Kumar et al., 2025; Oberemok et al., 2025a).

2.2. Comparison with RNAi and CRISPR/Cas

• CUADb is one of three antisense technologies alongside RNAi and CRISPR/Cas.
• All involve duplex formation with unmodified nucleic acids and nuclease-mediated gene silencing: RNAi: guide RNA–mRNA (Argonaute); CUADb: guide DNA–rRNA (DNA-guided rRNase); CRISPR/Cas: guide RNA–DNA (Cas proteins) (Kumar et al., 2025).

2.3. Use of DNAInsector for Oligo Design

• DNAInsector generates target oligonucleotides in ~10 seconds (DNAInsector, 2025).
• Simplifies oligo pesticide design, making it accessible even to non-specialists.

3. Practical Implementation and Application Strategy

3.1. Selection of Pest Targets

• 12 DNA pesticides developed for important pests (mostly Sternorrhyncha and 1 spider mite), chosen due to experimental precedence (Oberemok et al., 2024a).

3.2. Oligo Pesticide Application Methods and Dosages

• Application via cold fogger or backpack sprayer using 10–20 micron droplets.
• Dosage: Trees: 10–12 g oligo in 180–200 L water per hectare (400–500 trees); Bushes/grasses: 9–10 g in 180–200 L (Oberemok et al., 2025c).

3.3. Oligo Pesticide Application Methods and Dosages

• Pesticides are primarily contact-based, not systemic. Direct contact with pest integument is required (Oberemok et al., 2025c).

4. Case Studies: Proposed Oligonucleotide Pesticides

4.1. Selection of Insect Pests

• 12 pests from 5 continents, including hemipterans like Myzus persicae, Diaphorina citri, Planococcus citri, and the spider mite Panonychus ulmi.

4.2. Pest-Specific Oligo Design and Targets

• All target 18S or 28S rRNA using 11-nt antisense DNA sequences (Table 1).

4.3. Global Representation of Species

• Origin of pests illustrated in Figure 2.
• Species include those affecting apples, potatoes, citrus, coffee, and forest trees (e.g., Adelges tsugae, Paracoccus marginatus) (Soltis et al., 2014; Buzzetti et al., 2015; García Morales et al., 2016; Alba-Tercedor et al., 2021; Kar et al., 2023).

5. Advantages of the ‘Genetic Zipper’ Method

5.1. Cost and Ease of Synthesis

• CUADb is more cost-effective than RNAi and CRISPR/Cas technologies (Oberemok et al., 2024c; 2024d).

5.2. Safety and Environmental Benefits

• Low carbon footprint, biodegradable, safe for non-target species (Gal’chinsky et al., 2023; Oberemok et al., 2024b).

5.3. High Target Specificity

• Antisense oligos designed for specific rRNA sequences reduce off-target impacts (Gal’chinsky et al., 2024; Oberemok et al., 2025a).

5.4. Resistance Management Potential

• Novel mode of action may help prevent development of target-site resistance (Oberemok et al., 2024c).

5.5. Predictive Design Using Related Species

• Effectiveness can be predicted for untested species using phylogenetic proximity (Oberemok et al., 2024a; Gavrilova et al., 2025).

6. Advantages of the ‘Genetic Zipper’ Method

6.1. Limited Pest Coverage

• Proven mainly for hemipterans and spider mites; broader validation is needed (Gavrilova et al., 2025).

6.2. Off-Target and Specificity Concerns

• Requires careful design to avoid unintended effects (Kumar et al., 2025).

6.3. Delivery System Limitation

• Field delivery methods are still being optimized (Oberemok et al., 2025).

6.4. Risk of Resistance Evolution

• New resistance mechanisms may eventually emerge (Gal’chinsky et al., 2024).

6.5. Production and Cost Issue

• While cheaper than some RNA-based tools, it still needs specialized synthesis setups (Hemant et al., 2025).

7. Conclusion and Future Outlook

The “genetic zipper” method represents a groundbreaking step toward the personalization of pesticide development, transforming the traditionally complex and resource-intensive process into a fast, rule-based system accessible even to non-specialists. Through digital tools like the DNAInsector platform, researchers and practitioners can rapidly design pest-specific oligonucleotide insecticides, making real-time responses to emerging pest threats more feasible than ever before. This capability opens new opportunities for tailored pest control solutions in both research and agricultural settings, including localized outbreaks and resistant populations. In practical terms, the CUADb-based approach fits seamlessly into integrated pest management (IPM) strategies due to its high specificity, environmental safety, and ease of application. It complements existing pest control methods by enabling selective suppression of target pests without harming beneficial organisms, making it suitable for use in organic and ecologically sensitive farming systems. However, the successful integration of this technology into mainstream agriculture will depend heavily on clear regulatory pathways and strong public engagement. While CUADb-based pesticides are non-transgenic and biodegradable, many regulatory systems are still adapting to emerging nucleic acid technologies. Therefore, transparent scientific communication, involvement of independent research bodies, and public education will be essential to ensure both regulatory approval and societal acceptance. The future of CUAD biotechnology hinges not only on its scientific merit but also on its ability to gain trust as a safe, flexible, and sustainable tool for modern agriculture (Oberemok et al., 2024d).