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Related Concept Videos

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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Homologous Recombination02:31

Homologous Recombination

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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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Conservative Site-specific Recombination and Phase Variation02:53

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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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CRISPR and crRNAs02:53

CRISPR and crRNAs

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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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[Methods to Increase the Efficiency of Knock-in of a Construct Encoding the HIV-1 Fusion Inhibitor, MT-C34 Peptide, into the CXCR4 Locus in the CEM/R5 T Cell Line].

Molekuliarnaia biologiia·2024
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[Donor DNA Modification with Cas9 Targeting Sites Improves the Efficiency of MTC34 Knock-in into the CXCR4 Locus].

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Generation of LEPR Knockout Rabbits with CRISPR/CAS9 System.

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Related Experiment Video

Updated: Jul 6, 2025

CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.

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Efficient Editing of the CXCR4 Locus Using Cas9 Ribonucleoprotein Complexes Stabilized with Polyglutamic Acid.

D S Golubev1, D S Komkov1,2, M V Shepelev1

  • 1Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia.

Doklady Biological Sciences : Proceedings of the Academy of Sciences of the USSR, Biological Sciences Sections
|January 8, 2024
PubMed
Summary
This summary is machine-generated.

Improving CRISPR/Cas9 gene editing efficiency is crucial for treating diseases like HIV. This study enhanced CRISPR/Cas9 by modifying Cas9 protein and stabilizing complexes, boosting gene editing for HIV therapies.

Keywords:
CRISPR/Cas9CXCR4HIVNLSPGARNPgenome editing

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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

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Area of Science:

  • Molecular Biology
  • Gene Therapy
  • Immunology

Background:

  • CRISPR/Cas9 gene editing offers potential for treating human diseases.
  • Enhancing gene editing efficiency is critical for therapeutic applications.
  • The CXCR4 gene is a key target for HIV gene therapy.

Purpose of the Study:

  • To improve the efficiency of CRISPR/Cas9 gene editing.
  • To enhance the editing of the CXCR4 locus for HIV gene therapy.
  • To investigate methods for increasing gene knockout and knock-in levels.

Main Methods:

  • Modified the Cas9 protein by adding Nuclear Localization Signals (NLS).
  • Stabilized Cas9 and guide RNA ribonucleoprotein complexes using poly-L-glutamic acid.
  • Assessed editing efficiency in CEM/R5 T cell lines and primary CD4+ T lymphocytes.

Main Results:

  • Achieved a 1.8-fold increase in CXCR4 knockout in a T cell line.
  • Demonstrated a 2-fold increase in knock-in of the MT-C34 HIV-1 fusion peptide inhibitor in primary CD4+ T cells.
  • The modifications significantly enhanced gene editing outcomes.

Conclusions:

  • The developed approach effectively increases CRISPR/Cas9 gene editing efficiency.
  • This method holds promise for advancing gene therapy strategies for HIV infection.
  • Optimized gene editing techniques are vital for successful therapeutic interventions.