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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

208
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...
208
CRISPR01:59

CRISPR

52.8K
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: Sep 9, 2025

Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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A general genome editing strategy using CRISPR lipid nanoparticle spherical nucleic acids.

Zhenyu Han1, Chi Huang1, Taokun Luo1

  • 1Department of Chemistry and International Institute for Nanotechnology, Northwestern University, Evanston, IL 60208.

Proceedings of the National Academy of Sciences of the United States of America
|September 4, 2025
PubMed
Summary

Researchers developed CRISPR lipid nanoparticle-spherical nucleic acids (LNP-SNAs) for efficient gene editing. These novel LNP-SNAs show improved delivery, reduced toxicity, and enhanced CRISPR-Cas editing efficiency compared to traditional lipid nanoparticles.

Keywords:
CRISPRgenome editinglipid nanoparticlerepair templatespherical nucleic acid

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Gene Therapy

Background:

  • CRISPR-Cas systems offer therapeutic potential for genetic disorders and cancers.
  • Efficient delivery of CRISPR components (Cas proteins, guide RNAs, donor DNA) is a major challenge.
  • Existing delivery vehicles can exhibit immunogenicity and toxicity, limiting clinical application.

Purpose of the Study:

  • To develop a novel, efficient, and biocompatible delivery platform for CRISPR-Cas systems.
  • To enhance cellular uptake, reduce cytotoxicity, and improve gene editing efficiency.
  • To evaluate the performance of CRISPR lipid nanoparticle-spherical nucleic acids (LNP-SNAs) compared to traditional lipid nanoparticles (LNPs).

Main Methods:

  • Combination of lipid nanoparticle (LNP) and spherical nucleic acid (SNA) technologies.
  • Synthesis of CRISPR LNP-SNAs with a surface-bound DNA shell.
  • Assessment of cellular uptake, cytotoxicity, and gene transfection efficiency across multiple cell lines.
  • Quantification of insertion-deletion mutation frequencies and homology-directed repair (HDR) efficiency.

Main Results:

  • CRISPR LNP-SNAs demonstrated 2-3 fold higher cellular uptake and gene transfection efficiency than LNPs.
  • LNP-SNAs exhibited reduced cytotoxicity compared to LNPs.
  • Average insertion-deletion mutation frequencies were 2-3 fold higher with LNP-SNAs.
  • Homology-directed repair efficiency was significantly improved, reaching 21 ± 7% with LNP-SNAs versus 8 ± 4% with LNPs.

Conclusions:

  • CRISPR LNP-SNAs represent a highly efficient and biocompatible delivery platform for CRISPR-Cas gene editing.
  • The enhanced performance of LNP-SNAs addresses key challenges in CRISPR delivery, including efficiency and safety.
  • This versatile platform holds significant promise for advancing CRISPR-based genetic medicines and other gene therapies.