Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Somatic to iPS Cell Reprogramming01:29

Somatic to iPS Cell Reprogramming

2.6K
Reprogramming alters the gene expression in somatic cells, transforming them into induced pluripotent stem (iPS) cells over several generations. Scientists can reprogram cells by introducing genes for four transcription factors—Oct4, Sox2, Klf4, and c-Myc (OSKM) by viral or non-viral methods. These factors are also known as Yamanaka factors after Shinya Yamanaka, who first generated iPS cells using mouse skin cells. Yamanaka was awarded the Nobel Prize in Physiology or Medicine in 2012...
2.6K

You might also read

Related Articles

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

Sort by
Same author

Validated GMP banking of a new set of genomically stable induced pluripotent stem cell lines using single-cell passaging.

Cytotherapy·2026
Same author

Deep immune-phenotyping of HLA-homozygous iPS-cardiomyocytes by spectral flow cytometry.

Frontiers in immunology·2026
Same author

Improved cryopreservation of cardiomyocyte aggregates differentiated from GMP iPSC in a 3D culture format.

Scientific reports·2026
Same author

GMP-compliant iPS cell lines show widespread plasticity in a new set of differentiation workflows for cell replacement and cancer immunotherapy.

Stem cells translational medicine·2024
Same author

Transneuronal Delivery of Cytokines to Stimulate Mammalian Spinal Cord Regeneration.

Methods in molecular biology (Clifton, N.J.)·2023
Same author

Pathophysiological Mechanisms of Cardiac Dysfunction in Transgenic Mice with Viral Myocarditis.

Cells·2023

Related Experiment Video

Updated: Jan 18, 2026

Efficient Generation and Editing of Feeder-free IPSCs from Human Pancreatic Cells Using the CRISPR-Cas9 System
09:16

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

Published on: November 8, 2017

10.6K

Sequential factor delivery enables efficient workflow for universal gene editing in clinical grade iPS cells.

Thomas Berger1, Elitsa Borisova1, Anna Gamerschlag1

  • 1Catalent Duesseldorf GmbH, Berghausener Straße 98, 40764, Langenfeld, Germany.

Scientific Reports
|September 12, 2025
PubMed
Summary

We optimized CRISPR gene editing for human induced pluripotent stem cells (iPSCs), achieving over 30% knock-in efficiency for cell therapy applications. This virus-free method enhances safety and immune compatibility for therapeutic use.

More Related Videos

CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation
08:31

CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation

Published on: June 20, 2019

14.7K
Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor
07:28

Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor

Published on: February 1, 2015

20.5K

Related Experiment Videos

Last Updated: Jan 18, 2026

Efficient Generation and Editing of Feeder-free IPSCs from Human Pancreatic Cells Using the CRISPR-Cas9 System
09:16

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

Published on: November 8, 2017

10.6K
CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation
08:31

CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation

Published on: June 20, 2019

14.7K
Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor
07:28

Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor

Published on: February 1, 2015

20.5K

Area of Science:

  • Stem Cell Biology
  • Gene Editing Technologies
  • Cellular Therapy

Background:

  • Human induced pluripotent stem cells (iPSCs) show great promise for cell therapy.
  • CRISPR technology enables gene modifications in iPSCs for enhanced therapeutic features.
  • Current genetic manipulation of iPSCs, especially knock-ins, is inefficient and may conflict with Good Manufacturing Practice (GMP).

Purpose of the Study:

  • To optimize a CRISPR-based gene editing procedure for efficient knock-in generation in iPSCs.
  • To develop a GMP-compatible, virus-free method for genetic manipulation of iPSCs.
  • To create and validate iPSC lines with enhanced immune compatibility and safety features.

Main Methods:

  • Systematic optimization of Cas9 and Cas12a-based ribonucleoprotein (RNP) complexes for gene editing.
  • Sequential delivery of RNPs and donor plasmids for virus-free knock-in.
  • Creation and characterization of homozygous iPSC lines with HLA class I depletion and an inducible caspase-9 suicide gene.

Main Results:

  • Achieved over 30% knock-in efficiency for full-length transgenes using the optimized method.
  • Successfully generated homozygous iPSC lines lacking HLA class I and expressing a safety switch.
  • Validated genomic integrity, differentiation capability, and safety switch functionality in GMP iPSC lines.

Conclusions:

  • The improved gene editing methodology offers a flexible and efficient platform for iPSC modification.
  • This virus-free approach overcomes GMP limitations associated with current gene editing techniques.
  • The developed iPSC lines are suitable for basic research and hold potential for cell therapy applications.