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Updated: Feb 5, 2026
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Embryo Microinjection and Knockout Mutant Identification of CRISPR/Cas9 Genome-Edited Helicoverpa Armigera Hübner
Published on: July 1, 2021
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A genome editing vector that enables easy selection and identification of knockout cells
Akira Nagasaki1, Yoshio Kato1, Keiichi Meguro2
1Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan.
Plasmid
|September 10, 2018
Summary
A new CRISPR/Cas9 vector, pGedit, simplifies gene editing. This novel tool enhances efficiency for creating knockout cells, making genome editing more accessible for researchers.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- CRISPR/Cas9 is a key tool for gene disruption.
- Current genome editing vectors are often inefficient and laborious.
- Improved methods are needed for efficient knockout cell generation.
Purpose of the Study:
- To develop a novel, high-efficiency genome editing vector.
- To simplify and accelerate the process of creating gene knockout cell lines.
- To introduce the pGedit vector for enhanced CRISPR/Cas9 applications.
Main Methods:
- Construction of the pGedit vector, incorporating Cas9, sgRNA, and EGFP-Bsr selection marker.
- Utilizing the pGedit vector for targeted gene disruption in human (ACTB, ACTG1) and mouse (Nes) cells.
- Employing EGFP-Bsr for rapid selection and monitoring of edited cells.
Main Results:
- The pGedit vector demonstrated high efficiency in genome editing across multiple gene targets.
- Successfully generated knockout cell lines for human ACTB, ACTG1, and mouse Nes genes.
- EGFP-Bsr marker facilitated easy selection and confirmed vector non-integration.
Conclusions:
- The pGedit vector significantly streamlines the genome editing process.
- pGedit offers a more efficient and user-friendly approach to generating knockout cells.
- This novel vector facilitates broader application of CRISPR/Cas9 technology.
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