Engineered Microbial Population for Bee RNAi Delivery
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Solution Overview
Problem
Current methods for inducing RNAi in honey bees, such as injecting or feeding dsRNA, are costly, unstable, and unreliable, and do not effectively address bee colony losses due to viral pathogens and Varroa mite infestations.
Innovation Solution
Genetically engineering native bacteria from the honey bee microbiome, like Snodgrassella alvi, to express heterologous nucleic acids that produce dsRNA, which can be administered to the bees, providing a stable and cost-effective means to induce RNAi against specific gene products of pathogens and pests.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If dsRNA is injected or fed to honey bees to induce RNAi, then gene expression downregulation is achieved, but the method is costly and operationally complex
Solution Approach 1:
The patent uses engineered bacteria as an intermediary carrier to deliver dsRNA to bees. Instead of directly injecting or feeding dsRNA (which is costly and complex), the bacteria naturally colonize the bee gut and continuously produce dsRNA, simplifying administration while maintaining RNAi effectiveness
Solution Approach 2:
The engineered bacteria autonomously produce and deliver dsRNA within the bee gut without requiring repeated external administration. The bacteria self-replicate and continuously express dsRNA, providing sustained gene silencing without ongoing human intervention
2Reliability
If dsRNA is produced in the laboratory and administered to bees, then RNAi is induced, but the dsRNA is unstable and rapidly degrades in the environment
Solution Approach 1:
The engineered bacteria continuously synthesize fresh dsRNA within the protected environment of the bee gut, eliminating the stability problems of externally administered dsRNA. The bacteria serve as living factories that autonomously produce stable, sustained levels of dsRNA
Solution Approach 2:
The bacteria act as a protective intermediary that shields dsRNA from environmental degradation. By producing dsRNA inside the bee's gut rather than administering it externally, the system protects the fragile dsRNA from UV radiation, temperature fluctuations, and enzymatic degradation in the external environment
3Reliability
If dsRNA is injected into bees to induce RNAi, then gene expression is downregulated, but the injection process is traumatic and reduces bee welfare
Solution Approach 1:
The bacteria serve as a benign intermediary that delivers dsRNA through natural colonization of the gut rather than through traumatic injection. The bees ingest the bacteria through feeders, and the bacteria naturally establish themselves in the gut environment without causing physical harm
Solution Approach 2:
The patent replaces the mechanical injection system (syringes, needles, physical penetration) with a biological delivery system where bacteria are ingested and colonize naturally. This substitutes traumatic mechanical delivery with gentle oral administration followed by natural bacterial colonization
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach allows for continuous production and delivery of dsRNA within the bee gut, effectively downregulating target gene expression, reducing bee mortality and susceptibility to diseases like Colony Collapse Disorder and Varroa mite infestations, and maintaining bacterial persistence over the bee's lifetime.
Implementation Method 1
one or more bacteria genetically engineered to express at least one heterologous nucleic acid
Implementation Method 2
RNA interference (RNAi) is a powerful technique to specifically downregulate gene expression in many eukaryotes by providing dsRNA with sequence identity to eukaryotic genes
Data Source
AI summary
Provided herein are genetically engineered bacteria that are native to a host insect microbiome. Further provided are methods of inducing RNA interference in an insect, such as a bee, by administering the genetically engineered bacteria.