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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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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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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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RNA Editing02:23

RNA Editing

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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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Related Experiment Video

Updated: Feb 20, 2026

Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

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RNA editing with CRISPR-Cas13.

David B T Cox1,2,3,4,5,6, Jonathan S Gootenberg1,2,3,4,7, Omar O Abudayyeh1,2,3,4,6

  • 1Broad Institute of Massachusetts Institute of Technology (MIT) and Harvard, Cambridge, MA 02142, USA.

Science (New York, N.Y.)
|October 27, 2017
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel RNA editing platform called REPAIR, using CRISPR-Cas13 and ADAR2. This system precisely corrects disease-causing mutations at the RNA level in mammalian cells, offering therapeutic potential.

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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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Related Experiment Videos

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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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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
09:51

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Genetic Engineering

Background:

  • RNA editing offers a promising strategy for treating genetic diseases by correcting disease-related sequences.
  • Type VI CRISPR-Cas systems, specifically the Cas13 enzyme, are RNA-guided nucleases with potential for programmable RNA manipulation.

Purpose of the Study:

  • To engineer a Cas13 ortholog for robust RNA knockdown.
  • To demonstrate RNA editing in mammalian cells using catalytically inactive Cas13 (dCas13) to guide adenosine deaminase acting on RNA 2 (ADAR2).
  • To develop a versatile RNA editing platform for research and therapeutics.

Main Methods:

  • Profiling of type VI CRISPR-Cas systems.
  • Engineering of a catalytically inactive Cas13 (dCas13) variant.
  • Co-expression of dCas13 with adenosine deaminase acting on RNA 2 (ADAR2) for targeted adenosine-to-inosine (A-to-I) RNA editing.
  • Development of a high-specificity variant and a minimized system for viral delivery.

Main Results:

  • Demonstrated successful RNA editing in mammalian cells using the REPAIR system.
  • Showcased the ability to edit full-length transcripts with pathogenic mutations without strict sequence constraints.
  • Engineered a high-specificity variant and a compact system suitable for viral delivery.

Conclusions:

  • The REPAIR system represents a novel and programmable RNA editing platform.
  • This technology has broad applicability in basic research, therapeutic development for genetic diseases, and biotechnology.
  • REPAIR offers a flexible approach to correct RNA sequences, bypassing the need for DNA modification.