Embryo Microinjection for Transgenesis in <em>Drosophila</em>

Guang Chen; Xinyi Zhang; Si Xu; Xiaotao Zhou; Wen Xue; Xuelian Liu; Jialong Yan; Nana Zhang; Jiwu Wang

doi:10.3791/66679

Embryo Microinjection for Transgenesis in Drosophila

JoVE Journal
Genetics
Author Produced

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Genetics

Embryo Microinjection for Transgenesis in Drosophila

Embryo Microinjection for Transgenesis in Drosophila

Full Text

3,747 Views

05:32 min

June 7, 2024

DOI: 10.3791/66679-v

Guang Chen*^1,2, Xinyi Zhang*^1,3, Si Xu^1,4, Xiaotao Zhou¹, Wen Xue¹, Xuelian Liu¹, Jialong Yan¹, Nana Zhang¹, Jiwu Wang^1,2,4

¹Clinical Research Institute, The Affiliated Nanhua Hospital, Hengyang Medical School,University of South China, ²Department of Clinical Laboratory, The Affiliated Nanhua Hospital, Hengyang Medical School,University of South China, ³Institute of Biochemistry and Molecular Biology, Hengyang Medical School,University of South China, ⁴Department of Neurology, The Affiliated Nanhua Hospital, Hengyang Medical School,University of South China

Overview

This article presents an optimized protocol for embryo microinjection, a crucial step for creating transgenic flies using phiC31 integrase-mediated transgenesis in Drosophila.

Key Study Components

Area of Science

Neuroscience
Genetics
Developmental Biology

Background

Transgenesis in Drosophila is essential for studying gene function.
Embryo microinjection is a key technique for constructing transgenic flies.
The phiC31 integrase system is a widely used method for transgenesis.
Optimizing microinjection techniques can improve success rates in generating transgenic lines.

Purpose of Study

To provide a detailed protocol for embryo microinjection in Drosophila.
To enhance the efficiency of creating transgenic flies.
To demonstrate the use of phiC31 integrase in transgenesis.

Methods Used

Embryo microinjection protocol using phiC31 integrase.
Use of beveled injection needles to minimize cytoplasmic leakage.
Application of air pressure during needle grinding for optimal tip shape.
Detailed steps for preparing and executing the microinjection process.

Main Results

Successful demonstration of embryo microinjection technique.
Improved needle design led to reduced cytoplasmic leakage.
Protocol allows for efficient generation of transgenic Drosophila.
Application of air pressure during grinding enhanced needle performance.

Conclusions

The optimized protocol facilitates the creation of transgenic flies.
Utilizing phiC31 integrase can streamline transgenesis processes.
Future studies can build on this protocol for various genetic experiments.

Frequently Asked Questions

What is the phiC31 integrase system?

The phiC31 integrase system is a method used for integrating transgenes into the Drosophila genome efficiently.

Why is embryo microinjection important?

Embryo microinjection is crucial for introducing genetic material into developing embryos to create transgenic organisms.

How does the beveled needle design improve results?

Beveled needles reduce cytoplasmic leakage during injection, leading to higher success rates in transgenesis.

What are the main challenges in embryo microinjection?

Challenges include maintaining embryo viability, minimizing leakage, and achieving precise injection.

Can this protocol be applied to other organisms?

While this protocol is optimized for Drosophila, similar techniques may be adapted for other model organisms.

What tools are necessary for this protocol?

Essential tools include a microinjection setup, beveled needles, and a foot air pump with a pressure gauge.

This article takes the phiC31 integrase-mediated transgenesis in Drosophila as an example and presents an optimized protocol for embryo microinjection, a crucial step for creating transgenic flies.

Transgenesis in drosophila is essential for studying gene function at the organism level. Embryo microinjection is a crucial step for construction of transgenic flies. Here, we take the phiC31 integrase-mediated transgenesis in drosophila as an example, and present a detailed protocol for embryo microinjection for transgenesis in drosophila.

Injection needles whose tips beveled by a grinder to mask embryonic block, or less large cytoplasmic leakage. However, the grinding solution can enter the beveled tip of a needle during the grinding process as the grinding solution inside the tip does not naturally dry up quickly. Our protocol uses a foot air pump with a pressure gauge to apply air pressure to a needle while grinding its tip.

View the full transcript and gain access to thousands of scientific videos