Related Experiment Video
Updated: Jun 22, 2025
12:21
Generating Recombinant Avian Herpesvirus Vectors with CRISPR/Cas9 Gene Editing
Published on: January 7, 2019
13.3K
Probing Baculovirus Vector Gene Essentiality for Foreign Gene Expression Using a CRISPR-Cas9 System.
Madhuja Chakraborty1, Jacqueline Powichrowski1, Mark R Bruder1
1Department of Chemical Engineering, Faculty of Engineering, University of Waterloo, Waterloo, ON, Canada.
Methods in Molecular Biology (Clifton, N.J.)
|July 1, 2024
Summary
The baculovirus expression vector system (BEVS) offers advantages for producing biologics. A new CRISPR-Cas9 method probes the baculovirus genome to identify genes for enhancing foreign protein production.
Area of Science:
- Molecular Biology
- Biotechnology
- Virology
Background:
- The baculovirus expression vector system (BEVS) is a valuable tool for producing biologics due to its safety and ability to perform mammalian-like post-translational modifications (PTMs) in insect cells.
- Despite its advantages, industrial adoption of BEVS lags behind other systems, potentially due to a lack of genetic tools and co-production of the vector with the desired product.
- The baculovirus genome remains largely unexamined for genetic enhancements that could optimize foreign gene expression.
Purpose of the Study:
- To develop and present a CRISPR-Cas9-based transfection-infection assay for investigating the baculovirus genome.
- To identify essential and nonessential genes within the baculovirus genome that can be manipulated to enhance foreign gene expression.
- To provide a novel genetic tool for improving the BEVS as an industrial production platform.
Main Methods:
- Development of a CRISPR-Cas9 system for targeted genetic manipulation of the baculovirus genome.
- Establishment of a transfection-infection assay to evaluate the impact of gene modifications on baculovirus replication and foreign gene expression.
- Systematic probing of the baculovirus genome to identify genes crucial for viral replication or foreign protein yield.
Main Results:
- The study outlines a methodology for a CRISPR-Cas9-based assay applicable to baculovirus genome research.
- This approach allows for the systematic identification of genes that can be targeted to enhance foreign gene expression.
- The described method provides a foundation for future genetic engineering of baculovirus vectors.
Conclusions:
- The development of advanced genetic tools, such as CRISPR-Cas9, is crucial for unlocking the full potential of the BEVS.
- Probing the baculovirus genome can reveal targets for genetic modification to improve foreign gene expression and optimize the system for industrial applications.
- This work lays the groundwork for enhancing the BEVS through targeted genomic investigation and engineering.
Related Concept Videos
01:59CRISPR
50.5K
Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
50.5K
02:53CRISPR and crRNAs
17.0K
Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
17.0K