Determining the Role of Maternally-Expressed Genes in Early Development with Maternal Crispants

Cara E. Moravec; Gabriella C. Voit; Francisco Pelegri

doi:10.3791/63177

Determining the Role of Maternally-Expressed Genes in Early Development with Maternal Crispants

JoVE Journal
Genetics

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JoVE Journal Genetics

Determining the Role of Maternally-Expressed Genes in Early Development with Maternal Crispants

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10:08 min

December 21, 2021

DOI: 10.3791/63177-v

Cara E. Moravec¹, Gabriella C. Voit¹, Francisco Pelegri¹

¹Laboratory of Genetics,University of Wisconsin-Madison

Overview

This study presents a novel CRISPR-Cas9 technique to identify maternal-effect phenotypes in early development. By utilizing multiplexing guide RNAs, researchers can rapidly uncover the roles of maternally-expressed genes in a single generation.

Key Study Components

Area of Science

Neuroscience
Developmental Biology
Genetics

Background

The function of many maternally-expressed genes during early development remains largely unknown.
Understanding these genes is crucial for insights into early embryogenesis.
Maternal mRNA transcripts play a significant role in the development of gametes.
Identifying maternal effect genes can enhance our knowledge of developmental processes.

Purpose of Study

To develop a method for the rapid identification of maternal effect genes.
To investigate the role of these genes in early embryonic development.
To provide a protocol that can be utilized by researchers in the field.

Methods Used

Microinjection of embryos at the one-cell stage.
Multiplexing guide RNAs targeting specific maternal genes.
Assessment of somatic mutations in injected embryos.
Creation of guide RNA templates using specific oligonucleotides.

Main Results

Successful identification of novel maternal effect phenotypes.
Demonstrated the effectiveness of the CRISPR-Cas9 technique in a single generation.
Provided insights into the function of maternal mRNA transcripts.
Established a protocol for future research in maternal gene function.

Conclusions

The CRISPR-Cas9 technique is a powerful tool for studying maternal-effect genes.
This research enhances our understanding of early developmental processes.
Future studies can build on this protocol to explore additional maternal genes.

Frequently Asked Questions

What is the significance of maternal-effect genes?

Maternal-effect genes are crucial for early embryonic development, influencing various developmental processes.

How does the CRISPR-Cas9 technique work in this study?

The technique uses multiplexing guide RNAs to target specific maternal genes, allowing for rapid identification of their effects.

What stage of development is targeted for microinjection?

Microinjection is performed at the one-cell stage of embryonic development.

What are the expected outcomes of this research?

The research aims to identify novel maternal effect phenotypes and enhance understanding of maternal gene functions.

Can this method be applied to other species?

While this study focuses on a specific model, the method can potentially be adapted for use in other species.

What are the implications of understanding maternal mRNA transcripts?

Understanding these transcripts can provide insights into the mechanisms of early embryogenesis and developmental biology.

Early development is dependent on maternally-inherited products, and the role of many of these products is currently unknown. Herein, we described a protocol that uses CRISPR-Cas9 to identify maternal-effect phenotypes in a single generation.

The role of many maternally-expressed genes during early development is currently unknown. This maternal CRISPR technique allows for the rapid identification of maternal effect genes and their role in development. Multiplexing guide RNAs to a single gene, allows researchers to identify novel maternal effect phenotypes in a single generation.

This research is beneficial to understand the function of mRNA transcripts in the gametes, which are necessary for early embryogenesis. The key to this technique is to micro inject the embryos early in the one cell stage, and check to see of the majority of the guide RNAs can make somatic mutations in the injected embryos. To create a guide RNA template for each gene specific oligonucleotide, analyte to the constant oligonucleotide and fill in the overhangs with T4 DNA polymerase.

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