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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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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...
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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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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Chromatin Modification in iPS Cells01:32

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Chromatin modification alters gene expression; therefore, scientists can add histone-modifying enzymes, histone variants, and chromatin remodeling complexes to somatic cells to aid reprogramming into pluripotent stem (iPS) cells.
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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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Updated: Aug 19, 2025

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery
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Decorating chromatin for enhanced genome editing using CRISPR-Cas9.

Evelyn Chen1,2, Enrique Lin-Shiao1,2, Marena Trinidad1,2

  • 1Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720.

Proceedings of the National Academy of Sciences of the United States of America
|December 2, 2022
PubMed
Summary

Researchers enhanced CRISPR-Cas genome editing by fusing Cas9 with PRDM9, a chromatin remodeler. This fusion significantly increased precise edits via homology-directed repair (HDR) without raising off-target mutations.

Keywords:
CRISPRchromatinepigeneticsgenome editing

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Area of Science:

  • Molecular Biology
  • Genome Editing Technologies
  • Epigenetics and DNA Repair

Background:

  • CRISPR-associated (Cas) enzymes offer RNA-guided genome editing capabilities.
  • Homology-directed repair (HDR) enables precise DNA edits but is often inefficient compared to indel-forming end-joining pathways.

Purpose of the Study:

  • To investigate if fusing Cas9 with the chromatin remodeler PRDM9 can enhance HDR efficiency in human cells.
  • To determine if PRDM9-Cas9 fusion impacts the balance between HDR and indel formation during genome editing.

Main Methods:

  • Constructed a fusion protein linking Cas9 with PRDM9, a factor known to deposit H3K36me3 and H3K4me3 marks.
  • Assessed HDR efficiency and indel formation in multiple human cell lines using the PRDM9-Cas9 fusion compared to unmodified Cas9.
  • Evaluated off-target editing events to ensure specificity.

Main Results:

  • The PRDM9-Cas9 fusion protein localized to the Cas9 cut site and interacted with chromatin.
  • Observed a threefold increase in HDR efficiency and a fivefold increase in the HDR:indel ratio.
  • No significant increase in off-target genome editing was detected across tested cell lines.

Conclusions:

  • PRDM9 fusion enhances HDR efficiency in CRISPR-Cas genome editing by influencing chromatin states at the target locus.
  • This strategy effectively boosts precise editing outcomes while maintaining specificity, offering a method to improve CRISPR applications.
  • Highlights the role of chromatin modifications in directing DNA repair pathway choice during genome editing.