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CRISPR01:59

CRISPR

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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 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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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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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: Oct 4, 2025

Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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CRISPR-Cas9‒Based Genomic Engineering in Keratinocytes: From Technology to Application.

Jos P H Smits1, Luca D Meesters1, Berber G W Maste1

  • 1Department of Dermatology, Radboud University Medical Center (Radboudumc), Radboud Institute for Molecular Life Sciences (RIMLS), Nijmegen, The Netherlands.

JID Innovations : Skin Science From Molecules to Population Health
|February 11, 2022
PubMed
Summary

CRISPR-Cas9 genome editing in dermatology shows variable success. This review identifies key factors for optimizing CRISPR-Cas9 strategies in keratinocyte cell sources for better experimental and clinical outcomes.

Keywords:
3D, three-dimensionalAAV, adeno-associated virusCOL7, type VII collagenEB, epidermolysis bullosaHPV16, human papillomavirus type 16IV, ichthyosis vulgarisJEB, junctional epidermolysis bullosaKC, keratinocyteRDEB, recessive dystrophic epidermolysis bullosahPSC, Human pluripotent stem celliKC, induced keratinocyteiPSC, induced pluripotent stem cell

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Area of Science:

  • Genomic Medicine
  • Dermatology Research
  • Gene Editing Technologies

Background:

  • CRISPR-Cas9 is a powerful genome-editing tool but faces challenges in efficiency, cell viability, and clonal cell line generation.
  • These limitations hinder its widespread application in various scientific disciplines, including dermatology.

Purpose of the Study:

  • To review CRISPR-Cas9 applications in experimental dermatology.
  • To identify critical factors influencing successful genome editing in keratinocytes (KCs).
  • To provide guidance for future CRISPR-Cas9 implementation in dermatology.

Main Methods:

  • Comprehensive literature review of CRISPR-Cas9 studies in experimental dermatology.
  • Analysis of strategies employed across different keratinocyte cell sources.
  • Identification of common practices and success factors.

Main Results:

  • Most studies utilize immortalized keratinocytes for generating knockout cell lines.
  • Variable genome-editing efficiencies and low success rates are frequently reported.
  • Key factors for successful CRISPR-Cas9 strategies are identified across different KC sources.

Conclusions:

  • Optimizing CRISPR-Cas9 strategies requires careful consideration of KC source and experimental design.
  • Immortalized KCs are commonly used but may not be optimal for all applications.
  • This review offers critical insights to guide future fundamental and clinical applications of CRISPR-Cas9 in dermatology.