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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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Retroviruses02:33

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Retroviruses and retrotransposons both insert copies of their genetic elements into the genome of the host cell. Thus, the viral genes are passed on when the host genome is replicated or translated. A typical retroviral DNA sequence contains 3-4 genes that encode the different proteins required for its structural assembly and function as a molecular parasite. This DNA is transcribed into a single mRNA, which is very similar in structure to conventional mRNAs, i.e., it is capped at the 5’...
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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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Retrovirus Life Cycles01:10

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Retroviruses have a single-stranded RNA genome that undergoes a special form of replication. Once the retrovirus has entered the host cell, an enzyme called reverse transcriptase synthesizes double-stranded DNA from the retroviral RNA genome. This DNA copy of the genome is then integrated into the host’s genome inside the nucleus via an enzyme called integrase. Consequently, the retroviral genome is transcribed into RNA whenever the host’s genome is transcribed, allowing the...
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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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Retroviruses are RNA viruses that have been shown to cause cancers in diverse species, including chickens, mice, cats, and monkeys. The RNA genomes of these viruses are first reverse-transcribed into single and then double-stranded DNA (dsDNA) copies. This dsDNA called proviral DNA then integrates into the host genome. Subsequently, the host cell transcribes the proviral DNA in concert with the chromosomal DNA. This leads to the production of viral RNA and proteins that assemble at the host...
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Related Experiment Video

Updated: Feb 4, 2026

Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio
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Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio

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Transient Retrovirus-Based CRISPR/Cas9 All-in-One Particles for Efficient, Targeted Gene Knockout.

Yvonne Knopp1, Franziska K Geis1, Dirk Heckl2

  • 1Institute of Experimental Hematology, Hannover Medical School, Hannover 30625, Germany.

Molecular Therapy. Nucleic Acids
|October 15, 2018
PubMed
Summary
This summary is machine-generated.

Researchers developed non-integrating retrovirus particles for transient CRISPR/Cas9 delivery, enabling efficient gene knockout without long-term enzyme expression risks. This method avoids cytotoxicity and offers dose-controlled gene editing for therapeutic applications.

Keywords:
CRISPR/Cas9 all-in-one particlecytotoxicitygammaretroviral MS2 chimeragenome editingtargeted gene knockouttransient delivery

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Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9
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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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Last Updated: Feb 4, 2026

Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio
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Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio

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Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9
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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 Editing Technologies
  • Retroviral Vector Development

Background:

  • CRISPR/Cas9 is a powerful gene editing tool but long-term expression of its enzymes can cause cytotoxicity.
  • Existing transient expression methods for CRISPR/Cas9 may have limitations in efficiency and safety.
  • Developing safer and more efficient gene editing delivery systems is crucial for therapeutic applications.

Purpose of the Study:

  • To develop non-integrating retrovirus-based particles for transient CRISPR/Cas9 delivery.
  • To achieve targeted gene knockout with reduced cytotoxicity compared to constitutive expression.
  • To enable efficient and dose-controlled delivery of CRISPR/Cas9 components.

Main Methods:

  • Utilized gammaretroviral packaging machinery to create all-in-one retroviral particles.
  • Delivered Streptococcus pyogenes Cas9 (SpCas9) mRNA and single-guide RNA transcripts transiently.
  • Assessed gene disruption efficiency in various cell types, including primary cells.

Main Results:

  • Efficient disruption of a surrogate reporter gene and endogenous genes (CXCR4, TP53) was achieved.
  • Transient particle knockout efficiencies reached 52%-80% of integrating vector efficiencies.
  • Transient SpCas9 delivery prevented cytotoxicity and cell cycle arrest observed with stable overexpression.

Conclusions:

  • Non-integrating retrovirus-based particles offer an efficient method for transient CRISPR/Cas9 delivery.
  • This approach mitigates risks associated with long-term DNA-modifying enzyme expression.
  • The developed system allows for dose-controlled gene editing with potential clinical relevance.