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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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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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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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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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Updated: Sep 28, 2025

CRISPR-Cas9-based Genome Engineering to Generate Jurkat Reporter Models for HIV-1 Infection with Selected Proviral Integration Sites
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A combinatorial CRISPR-Cas12a attack on HIV DNA.

Minghui Fan1, Ben Berkhout1, Elena Herrera-Carrillo1

  • 1Laboratory of Experimental Virology, Department of Medical Microbiology, Amsterdam UMC, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.

Molecular Therapy. Methods & Clinical Development
|March 31, 2022
PubMed
Summary
This summary is machine-generated.

Dual CRISPR-Cas12a RNA (crRNA) therapy shows enhanced antiviral activity against HIV, preventing viral escape and inactivating the virus through hypermutation in cell cultures.

Keywords:
CRISPR-Cas12aHIV-1curedual crRNAslentiviral vector

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Area of Science:

  • Molecular Biology
  • Gene Editing Technologies
  • Virology

Background:

  • CRISPR-Cas12a is a gene editing tool with potential for reduced off-target effects compared to Cas9.
  • Previous studies showed Cas12a with single crRNA could neutralize HIV in cell culture.
  • HIV can develop resistance to single crRNA by mutating target sequences.

Purpose of the Study:

  • To evaluate the antiviral efficacy of dual CRISPR-Cas12a crRNA combinations against HIV.
  • To investigate HIV's escape mechanisms and resistance development under dual crRNA therapy.
  • To confirm the absence of replication-competent HIV after treatment.

Main Methods:

  • Testing seven dual crRNA combinations against HIV in infected T cell lines.
  • Analyzing HIV proviral genomes for mutations at Cas12a target sites.
  • Assessing viral inactivation mechanisms, including hypermutation and sequence excision.

Main Results:

  • Dual crRNA combinations demonstrated more robust antiviral activity than single crRNA.
  • The dual-crRNA therapy effectively prevented HIV escape in long-term cultures.
  • HIV inactivation occurred primarily through hypermutation (indel mutations) rather than excision.

Conclusions:

  • Dual CRISPR-Cas12a crRNA therapy offers a more effective strategy against HIV than single crRNA.
  • This approach prevents viral escape and leads to HIV inactivation via cellular DNA repair mechanisms.
  • The findings support the potential of dual crRNA therapy for durable HIV control.