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Related Concept Videos

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...
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 Short...

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Related Experiment Video

Updated: May 14, 2026

Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins
07:20

Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins

Published on: June 14, 2018

Reprogramming endogenous NK circuits by highly efficient nonviral genome editing.

Rih-Sheng Huang1,2,3, Shee Kwan Phung1,2, Darin Sumstad2,4

  • 1Division of Hematology, Oncology and Transplantation, Department of Medicine, Medical School, University of Minnesota, Minneapolis, MN, USA.

The Journal of Experimental Medicine
|May 13, 2026
PubMed
Summary
This summary is machine-generated.

This study presents a novel nonviral engineering platform for natural killer (NK) cells, achieving high homology-directed repair (HDR) efficiency for immunotherapy development. The platform enables precise genetic modifications, enhancing NK cell function and manufacturing scalability.

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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

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Related Experiment Videos

Last Updated: May 14, 2026

Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins
07:20

Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins

Published on: June 14, 2018

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery
07:49

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery

Published on: May 30, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
09:51

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

Area of Science:

  • Immunology
  • Cell Biology
  • Biotechnology

Background:

  • Natural killer (NK) cells are potent immunotherapy agents but face limitations in nonviral genetic engineering.
  • Challenges include low homology-directed repair (HDR) efficiency, DNA toxicity, and manufacturing complexities.
  • Existing methods hinder precise and scalable engineering of NK cell effector functions.

Purpose of the Study:

  • To develop a high-yield, nonviral knock-in platform for NK cell engineering.
  • To overcome limitations in precision engineering, DNA toxicity, and manufacturing.
  • To enable programmable editing of NK cell functions for therapeutic applications.

Main Methods:

  • Extensive rational screening to optimize homology-directed repair (HDR) insertion.
  • Hijacking endogenous transcriptional programs to install genetic circuits at defined genomic loci.
  • Integration of synthetic circuits (positive feedback, hypoxia-responsive) for context-dependent NK cell responses.

Main Results:

  • Achieved approximately 90% HDR insertion efficiency with 100% post-editing recovery.
  • Engineered NK cells with enhanced persistence and dual CAR expression via a CISH locus circuit.
  • Restored NK cell cytotoxicity under hypoxia using a PFKFB4-gated IL-12 circuit.
  • Demonstrated compatibility with Good Manufacturing Practice (GMP) and clinical-scale expansion.

Conclusions:

  • The developed nonviral platform offers a scalable framework for precise NK cell engineering.
  • Programmable editing of NK cell effector functions is achievable for therapeutic and research use.
  • This approach addresses key limitations in current NK cell-based immunotherapy development.