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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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CRISPR01:59

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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 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.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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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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Related Experiment Video

Updated: Nov 16, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Efficient Immune Cell Genome Engineering with Enhanced CRISPR Editing Tools.

Waipan Chan1, Rachel A Gottschalk2, Yikun Yao3

  • 1Lymphocyte Biology Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; waipan.chan@nih.gov rgermain@niaid.nih.gov.

Immunohorizons
|February 24, 2021
PubMed
Summary
This summary is machine-generated.

New CRISPR tools enhance genome editing in immune cells like macrophages and lymphocytes. These methods overcome previous challenges, offering efficient alternatives for both mouse and human cell engineering.

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Area of Science:

  • Immunology
  • Molecular Biology
  • Genome Engineering

Background:

  • CRISPR technology has transformed genome engineering but faces challenges in modifying immune cells.
  • Innate immune cells (macrophages) and adaptive immune cells (lymphocytes) are difficult to edit due to cell pathology, low targeting efficiency, and issues with donor template delivery.

Purpose of the Study:

  • To develop novel CRISPR-based genome editing strategies for efficient modification of immune cells.
  • To overcome limitations of existing CRISPR methods in targeting macrophages and lymphocytes.

Main Methods:

  • Developed three alternative CRISPR genome editing methods utilizing a hit-and-run transient expression strategy.
  • Integrated a CRISPR activation (CRISPRa) system for enhanced gene expression.
  • Applied strategies to both murine and human immune cell types.

Main Results:

  • Achieved efficient CRISPR-based genome editing in challenging immune cell populations.
  • Demonstrated successful application in both innate (macrophages) and adaptive (lymphocytes) immune cells.
  • Provided effective alternatives to existing transient transfection methods for donor template delivery.

Conclusions:

  • The developed CRISPR tools and strategies offer efficient genome engineering solutions for immune cells.
  • These methods are applicable to a broad range of hematopoietic and other mammalian cell types.
  • Represents a significant advancement in the study of gene-phenotype relationships in the immune system.