Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

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...

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Carbon dot-based polyplexes with cell penetration peptides for gene transfection.

RSC advances·2025

Same author

A Physiologically Based Pharmacokinetic (PBPK) Study to Assess the Adjuvanticity of Three Peptides in an Oral Vaccine.

Pharmaceutics·2024

Same author

Cell-Penetrating Peptides: Promising Therapeutics and Drug-Delivery Systems for Neurodegenerative Diseases.

Molecular pharmaceutics·2024

Same author

A Method for Using Cell-Penetrating Peptides for Loading Plasmid DNA into Secreted Extracellular Vesicles.

Biomolecules·2023

Same author

Aggregation Limiting Cell-Penetrating Peptides Derived from Protein Signal Sequences.

International journal of molecular sciences·2023

Same author

In Silico Studies to Support Vaccine Development.

Pharmaceutics·2023

Same journal

Mapping the 3D Chromosome Organization of a Biosynthetic Gene Cluster by Capture Hi-C (CHi-C).

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Mapping the 3D Chromosome Organization of Streptomyces by Hi-C.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

CUT&Tag Epigenomic Profiling of Biosynthetic Gene Clusters in Arabidopsis thaliana.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Rhizobium rhizogenes-Mediated Hairy Root Transformation Protocol for Lotus japonicus and Other Legumes.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Characterization of Bioactive Saponins from Sea Cucumbers.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Methods for Functional Validation of Terpenoid Metabolic Clusters in Nicotiana benthamiana and Aspergillus oryzae.

Methods in molecular biology (Clifton, N.J.)·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Oct 13, 2025

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution

Published on: February 26, 2019

CRISPR/Cas9 Plasmid Delivery Through the CPP: PepFect14.

Luca Falato¹, Birgit Vunk², Ülo Langel^2,3

¹Department of Biochemistry and Biophysics, Arrhenius Laboratories for Natural Sciences, Stockholm University, Stockholm, Sweden. luca.falato92@gmail.com.

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

|November 12, 2021

Summary

This summary is machine-generated.

This study introduces a novel gene editing method using CRISPR-Cas9 and Cell Penetrating Peptide (CPP) PepFect14 for efficient gene knockdown and expression in mammalian cells. The technique offers simultaneous readouts for research applications.

Keywords:

CPP CRISPR Gene editing Luciferase activity PepFect14 Peptide

More Related Videos

Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9

Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9

Published on: August 25, 2018

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Published on: October 18, 2022

Related Experiment Videos

Last Updated: Oct 13, 2025

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution

Published on: February 26, 2019

Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9

Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9

Published on: August 25, 2018

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Published on: October 18, 2022

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

Gene editing technologies are crucial for biological research.
CRISPR-Cas9 is a powerful tool for targeted genome modification.
Efficient delivery of gene editing components into cells is a key challenge.

Purpose of the Study:

To develop a novel protocol for gene editing using CRISPR-Cas9.
To utilize Cell Penetrating Peptide (CPP) PepFect14 for enhanced plasmid delivery.
To achieve simultaneous gene knockdown and reporter gene expression in mammalian cells.

Main Methods:

CRISPR-Cas9 plasmid encoding for gene knockdown and Green Fluorescent Protein (GFP) was complexed with PepFect14.
The pDNA/CPP complex was delivered into a Bomirsky Hamster Melanoma pLuc cell line.
Luciferase gene knockdown and GFP expression were assessed as readouts.
Cy5-labeled plasmid was used for visual confirmation of cellular uptake via fluorescent microscopy.

Main Results:

Successful delivery of the CRISPR-Cas9 plasmid into the target cell line using PepFect14.
Demonstrated knockdown of the luciferase gene.
Confirmed expression of the Green Fluorescent Protein (GFP).
Visual confirmation of pDNA/CPP complex cellular uptake was achieved.

Conclusions:

The developed protocol enables efficient gene editing in mammalian cells.
PepFect14 serves as an effective delivery vehicle for CRISPR-Cas9 plasmids.
The system provides simultaneous, easily measurable readouts for gene editing efficiency.