CRISPR/Cas9 in insects: Applications, best practices and biosafety concerns

Area of Science:

• Molecular Biology
• Genetics
• Entomology

Background:

• CRISPR/Cas9, initially a bacterial immune system, is now a versatile genome editing tool.
• Its high specificity and broad applicability have led to its use in diverse organisms, including insects, for research and applied purposes.
• CRISPR applications in insects are rapidly expanding for gene function studies and modifying economically important species.

Purpose of the Study:

• To review current successes of CRISPR/Cas9 applications in various insects and insect cells.
• To summarize best practices for CRISPR/Cas9 experimental setup in insects for researchers.
• To discuss the biosafety implications of releasing genetically edited insects into the environment.

Main Methods:

• Literature review of published studies on CRISPR/Cas9 in insect genome editing.
• Synthesis of experimental parameters influencing CRISPR/Cas9 efficacy, including Cas9/guide RNA forms, delivery methods, and target gene selection.
• Analysis of off-target effects and their implications.

Main Results:

• CRISPR/Cas9 has demonstrated success across a wide range of insect species and cell types.
• Variability in experimental outcomes is attributed to differences in specific parameters like reagent types and delivery systems.
• Identification of key factors influencing successful gene editing in insects.

Conclusions:

• CRISPR/Cas9 is a transformative technology for insect science, enabling both fundamental and applied research.
• Standardizing experimental parameters and addressing off-target effects are crucial for reproducible results.
• Careful consideration of biosafety is essential before environmental release of CRISPR/Cas9-edited insects.