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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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CRISPR/Cas9 Genome Editing01:28

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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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Homologous Recombination02:31

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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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CRISPR and crRNAs02:53

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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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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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Updated: Sep 20, 2025

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

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Harnessing CRISPR-Cas9 for Epigenetic Engineering.

Rosa S Guerra-Resendez1, Isaac B Hilton2,3,4

  • 1Systems, Synthetic, and Physical Biology PhD Program, Rice University, Houston, TX, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 6, 2022
PubMed
Summary
This summary is machine-generated.

This study outlines a method for epigenome editing using CRISPR/dCas9 technology. It details steps from guide RNA design to validating epigenetic changes via chromatin immunoprecipitation (ChIP).

Keywords:
CRISPRDeactivated Cas9Epigenome editingGuide RNASynthetic biology

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Area of Science:

  • Molecular Biology
  • Genetics
  • Epigenetics

Background:

  • Epigenome editing enhances precision by linking epigenetic effectors to dCas9.
  • Targeting regulatory DNA regions like promoters and enhancers is key.

Purpose of the Study:

  • To describe a methodology for epigenome editing experiments.
  • To provide a step-by-step guide for researchers.

Main Methods:

  • Guide RNA (gRNA) design and cloning.
  • Transient transfection of gRNA and CRISPR/dCas9-based epigenetic effector.
  • Chromatin immunoprecipitation (ChIP) for validation.

Main Results:

  • Successful implementation of the described epigenome editing workflow.
  • Validation of targeted epigenetic state modifications.

Conclusions:

  • The described methodology provides a foundational approach for epigenome editing.
  • This technique allows for precise modification of epigenetic states at specific genomic loci.