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CRISPR

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...

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Related Experiment Video

Updated: May 23, 2025

Efficient Generation and Editing of Feeder-free IPSCs from Human Pancreatic Cells Using the CRISPR-Cas9 System

Efficient Generation and Editing of Feeder-free IPSCs from Human Pancreatic Cells Using the CRISPR-Cas9 System

Published on: November 8, 2017

Targeted Human Editing Using CRISPR Technology in Neural Stem Cells.

Alvaro Gutierrez-Uzquiza^1,2, Paloma Bragado^3,4

¹Department of Biochemistry and Molecular Biology, Pharmacy Faculty, Complutense University of Madrid, Madrid, Spain. alguuz@ucm.es.

Methods in Molecular Biology (Clifton, N.J.)

|March 11, 2025

Summary

This summary is machine-generated.

This study introduces a lentiviral protocol for CRISPR-Cas9 gene editing in mammalian cells. This method facilitates the study of gene function by enabling precise DNA modifications for research applications.

Keywords:

CRISPR/Cas9 Gene editing Lentivirus Neural stem cells

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Last Updated: May 23, 2025

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Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

CRISPR-Cas9 is a powerful gene-editing tool utilizing guide RNA and the Cas9 enzyme to induce targeted DNA double-strand breaks.
These breaks are repaired via nonhomologous end joining (NHEJ) for mutations or homology-directed repair (HDR) for precise edits.
Efficient delivery of CRISPR components is critical for successful genome editing applications, including the generation of knockout models.

Purpose of the Study:

To present a robust protocol for CRISPR-Cas9 gene editing in mammalian cells.
To leverage lentiviral vectors for effective delivery of the CRISPR-Cas9 system.
To facilitate the study of gene function through precise genome modification.

Main Methods:

Development of a lentiviral-based delivery system for CRISPR-Cas9 components.
Application of the protocol to mammalian cell lines for gene editing.
Utilizing guide RNA to direct Cas9 nuclease activity to specific genomic loci.

Main Results:

Successful generation of targeted gene edits in mammalian cells via CRISPR-Cas9.
Demonstration of lentiviral vectors as an effective delivery method for CRISPR components across various cell types.
Facilitation of gene function studies through reliable genome editing.

Conclusions:

The lentiviral delivery protocol provides a valuable method for CRISPR-Cas9-mediated genome editing in mammalian cells.
This approach enhances the study of gene function in both in vitro and in vivo models.
The protocol supports diverse research applications in basic science, biotechnology, and biomedical research.