Related Experiment Video

Updated: Mar 22, 2026

Using CRISPR/Cas9 Gene Editing to Investigate the Oncogenic Activity of Mutant Calreticulin in Cytokine Dependent Hematopoietic Cells
10:21

Using CRISPR/Cas9 Gene Editing to Investigate the Oncogenic Activity of Mutant Calreticulin in Cytokine Dependent Hematopoietic Cells

Published on: January 5, 2018

13.8K

Using CRISPR/Cas9 Technology for Manipulating Cell Death Regulators

Andrew J Kueh1,2, Marco J Herold3,4

  • 1Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, VIC, 3052, Australia.

Methods in Molecular Biology (Clifton, N.J.)
|April 25, 2016
PubMed

Insights

This study introduces an inducible lentiviral vector for Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 gene editing. This method improves targeting efficiency and reduces lethality in essential genes.

Area of Science:

  • Molecular Biology
  • Gene Editing Technologies
  • Biotechnology

Background:

  • CRISPR/Cas9 is a powerful gene editing tool for creating targeted mutations.
  • Constitutive CRISPR/Cas9 expression can lead to low efficiency and lethality from essential gene mutations.

Purpose of the Study:

  • To develop an inducible CRISPR/Cas9 system for enhanced gene editing.
  • To overcome limitations of constitutive CRISPR/Cas9 expression.

Main Methods:

  • Utilized a lentiviral vector platform for inducible CRISPR/Cas9 delivery.
  • Employed cell transduction and enrichment strategies.
  • Induced CRISPR/Cas9 expression for targeted mutations.

Main Results:

  • Achieved rapid transduction and enrichment of CRISPR/Cas9 positive cells.
  • Demonstrated high levels of targeted mutations upon induction.
  • Showcased improved efficiency compared to constitutive expression.

Conclusions:

  • An inducible lentiviral vector platform enhances CRISPR/Cas9 gene editing efficiency.
  • This inducible system mitigates issues of low targeting efficiency and lethality.
  • Offers a more controlled and effective approach for CRISPR/Cas9 applications.
Keywords:
CRISPR/Cas9Guide RNAInducibleLentiviral

More Related Videos

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization08:20

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization

Published on: September 2, 2021

4.7K
Electroporation-Based CRISPR-Cas9-Mediated Gene Knockout in THP-1 Cells and Single-Cell Clone Isolation09:29

Electroporation-Based CRISPR-Cas9-Mediated Gene Knockout in THP-1 Cells and Single-Cell Clone Isolation

Published on: February 28, 2025

3.9K

Related Concept Videos

CRISPR01:59

CRISPR

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...
58.9K
CRISPR01:59

CRISPR

18.8K
CRISPR/Cas9 Genome Editing01:28

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...
2.5K