Pupal and Adult Injections for RNAi and CRISPR Gene Editing in <em>Nasonia vitripennis</em>

Elena Dalla Benetta; Duverney Chaverra-Rodriguez; Jason L. Rasgon; Omar S. Akbari

doi:10.3791/61892

ナソニアビトリペニスにおけるRNAiおよびCRISPR遺伝子編集のためのプパルおよび成人注射

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Pupal and Adult Injections for RNAi and CRISPR Gene Editing in Nasonia vitripennis

ナソニアビトリペニスにおけるRNAiおよびCRISPR遺伝子編集のためのプパルおよび成人注射

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08:41 min

December 4, 2020

DOI: 10.3791/61892-v

Elena Dalla Benetta*¹, Duverney Chaverra-Rodriguez*¹, Jason L. Rasgon², Omar S. Akbari¹

¹Division of Biological Sciences, Section of Cell and Developmental Biology,University of California, San Diego, ²Department of Entomology, The Center for Infectious Disease Dynamics, The Huck Institutes of the Life Sciences,The Pennsylvania State University

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Overview

This study presents a method for efficient pupal and adult injections in Nasonia vitripennis, providing a practical alternative to traditional embryonic microinjection. The technique enables functional analysis of genes through RNA interference (RNAi) and CRISPR/Cas9 genome editing.

Key Study Components

Research Area

Genetic engineering in non-canonical insect models
Functional genetics of Nasonia vitripennis
Applications of RNAi and CRISPR/Cas9 techniques

Background

Challenges in embryonic microinjection for parasitoid wasps
The need for accessible methods in genetic studies
Significance of using Nasonia as a model organism

Methods Used

Pupal and adult injection techniques for gene manipulation
Nasonia vitripennis as the biological system
Utilization of RNAi and CRISPR/Cas9 technologies

Main Results

Successful implementation of the injection method with high survival rates (15-100%)
Observation of desired phenotypes through gene editing
Verification of the effectiveness of RNAi and CRISPR without complicated procedures

Conclusions

The method demonstrates a significant advancement in the field of insect genetics
It opens new avenues for functional genetic studies in various insect species

Frequently Asked Questions

What is the purpose of this study?

The study aims to introduce a simpler approach for genetic manipulation in Nasonia vitripennis compared to traditional embryonic microinjections.

What are the two main techniques used in this research?

The techniques utilized are RNA interference (RNAi) and CRISPR/Cas9 genome editing.

Why is Nasonia vitripennis chosen as a model organism?

Nasonia vitripennis is a non-canonical insect model that allows researchers to investigate genetic mechanisms in parasitoid wasps.

What are the benefits of using pupal and adult injections?

These methods are faster, require less specialized equipment, and allow for heritable germline mutations compared to embryo injections.

What is the significance of achieving high survival rates?

High survival rates indicate the effectiveness and reliability of the injection method for genetic studies.

Can this method be applied to other insect species?

Yes, the optimized method may also facilitate functional studies in other insect species beyond Nasonia.

How does this study contribute to the field of genetics?

It provides accessible methods for genetic modification, expanding research possibilities in developmental biology and molecular genetics.

ここでは、 ナソニアビトリペンシス での効率的なプパルおよび成人注射を胚微小注射の代替手段として説明し、RNA干渉(RNAi)またはCRISPR/Cas9ゲノム編集によるRNAサイレンシングまたは遺伝子ノックアウトのいずれかを使用して目的の遺伝子の機能解析を可能にする。

寄生バチのような非標準的な昆虫モデルの研究は、胚マイクロインジェクションが非常に困難であるため、限られています。私たちの方法はこの問題を回避し、部位特異的および遺伝性の生殖細胞変異を可能にします。蛹や成体の女性への注射は迅速で、胚マイクロインジェクションを成功させるために必要な高価な機器や専門的なトレーニングの必要性を排除します。

この技術により、高価な機器にアクセスできない専門の研究室では、モデルとして使用している昆虫種の機能研究を行うことができなくなります。最適化後、この方法は、Nasoniaの他の多くの遺伝子の機能遺伝学を研究するために使用できます。また、この方法は、ナソニアや他の昆虫の遺伝子形質転換のための核酸送達にも利用できると考えています。

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