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CRISPR/Cas9 Genome Editing

CRISPR/Cas9 Genome Editing

The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...

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Updated: Mar 8, 2026

Genome Engineering of Primary Human B Cells Using CRISPR/Cas9

Genome Engineering of Primary Human B Cells Using CRISPR/Cas9

Published on: November 3, 2020

Baculovirus-based genome editing in primary cells.

Maysam Mansouri¹, Zahra Ehsaei², Verdon Taylor²

¹Paul Scherrer Institute, Biomolecular Research, Molecular Cell Biology, CH-5232 Villigen, Switzerland; ETH Zürich, Department of Biology, CH-8093 Zürich, Switzerland.

|January 26, 2017

Summary

This summary is machine-generated.

This study introduces new baculovirus-based plasmids for simplified genome editing using CRISPR/Cas9. These tools efficiently facilitate gene editing in primary cells and induced pluripotent stem cells via Homology Directed Repair.

Keywords:

Baculovirus CRISPR/Cas9 Genome editing Primary cells

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Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

CRISPR/Cas9 technology enables precise DNA modification by inducing double-strand breaks.
Homology Directed Repair (HDR) requires simultaneous delivery of repair templates alongside nucleases and guide RNAs.
Existing viral vectors like lentivirus and adenovirus have limited capacity for large HDR constructs.

Purpose of the Study:

To present novel Acceptor plasmids for the MultiPrime baculovirus system.
To simplify the cloning process for baculovirus-mediated genome editing.
To demonstrate the efficacy of the enhanced MultiPrime system in primary cells and induced pluripotent stem cells (iPS).

Main Methods:

Development and utilization of new Acceptor plasmids for the MultiPrime baculovirus system.
Genome editing experiments in primary cells with limited lifespan.
Genome editing experiments in induced pluripotent stem cells (iPS).

Main Results:

The new Acceptor plasmids streamline the construction of baculoviruses for genome editing.
The MultiPrime system, enhanced with new plasmids, successfully performed genome editing in primary cells.
The system demonstrated functionality in induced pluripotent stem cells (iPS), enabling precise genetic modifications.

Conclusions:

The improved MultiPrime baculovirus system offers a simplified and efficient method for genome editing.
This approach overcomes the capacity limitations of other viral vectors for delivering large HDR constructs.
The developed plasmids facilitate advanced genome engineering applications in various cell types, including stem cells.