Temperature-Controlled Insect Suppression Using Cpf1 Nuclease
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for generating sterile insect populations for control and modification are labor-intensive and face scalability issues during commercialization, relying on labor-intensive selection of transgenic males and females for genetic crosses.
Innovation Solution
The use of a temperature-dependent RNA-guided nuclease, Cpf1 (Cas12a), which allows for the generation of single insect lines containing both a nuclease and gRNA-expressing transgenes that are inactive at lower temperatures, enabling the production of sterile males when switched to higher temperatures, thus simplifying the scale-up process.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If labor-intensive selection of transgenic males and females is used to generate genetic crosses, then sterile male insects can be produced, but the process becomes unsuitable for scale-up during commercialization
Solution Approach 1:
The patent applies parameter changes by utilizing temperature as a control parameter to regulate nuclease activity. At permissive temperatures, the nuclease is inactive and allows normal insect development. At non-permissive temperatures, the nuclease becomes active and produces sterile males. This temperature-dependent parameter change enables automatic differentiation without labor-intensive selection processes, making the system scalable for commercialization.
Solution Approach 2:
The patent replaces the mechanical/labor-intensive selection system with a temperature-based biological control system. Instead of manually selecting and crossing specific transgenic individuals, the system uses temperature-induced nuclease activation to automatically generate sterile males, eliminating the need for labor-intensive genetic crossing operations while maintaining reliable production.
2Stability of the object's composition
If transgenic lines are maintained at lower temperatures, then the lines remain stable and fertile, but sterile male production is prevented
Solution Approach 1:
The patent applies dynamics by making the nuclease activity temperature-dependent rather than static. The same transgenic line can dynamically switch between two states: at permissive temperatures, the nuclease is inactive and the line remains stable and fertile for maintenance; at non-permissive temperatures, the nuclease becomes active and generates sterile males. This dynamic response to temperature changes allows the system to adapt between line maintenance and sterile male production modes.
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 efficient generation of sterile males without the need for labor-intensive selection, enabling temperature-controlled insect population engineering and suppression systems, and can be used to time gene drive activity for optimal effectiveness in the wild.
Implementation Method 1
the use of an RNA-guided nuclease, such as Cpf1 (aka Cas12a) which has a temperature-dependent activity
Data Source
AI summary
The present disclosure provides methods for generating insect strains that can be used for genetic suppression or modification of insect populations. The method of the present disclosure covers both gene drive (GD) approaches and genetic sterile insect technique (gSIT, aka pgSIT), and uses an RNA-guided nuclease, such as Cpf1 (aka Cas12a), which has a temperature-dependent activity and turns genetic approaches for insect control to temperature-controllable systems.