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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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RNA Editing02:23

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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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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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Experimental RNAi02:15

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RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
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Related Experiment Video

Updated: Aug 9, 2025

A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
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A Split CRISPR/Cas13b System for Conditional RNA Regulation and Editing.

Ying Xu1, Na Tian1, Huaxia Shi1

  • 1Department of Chemistry, Case Western Reserve University, 2080 Adelbert Road, Cleveland, Ohio 44106, United States.

Journal of the American Chemical Society
|February 22, 2023
PubMed
Summary
This summary is machine-generated.

Researchers developed a new CRISPR/Cas13b system controllable by abscisic acid (ABA) and light. This system precisely regulates RNA levels and modifications, advancing RNA research tools.

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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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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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Area of Science:

  • Molecular Biology
  • Biotechnology
  • RNA Biology

Background:

  • The CRISPR/Cas13b system is a powerful tool for RNA studies.
  • Precise control over Cas13b activity is crucial for understanding RNA functions.
  • Existing tools often interfere with native RNA activities.

Purpose of the Study:

  • To engineer a split Cas13b system for conditional RNA regulation.
  • To develop an ABA-inducible system for precise RNA manipulation.
  • To explore light-based control of RNA modification deposition.

Main Methods:

  • Engineering a split CRISPR/Cas13b system activated by abscisic acid (ABA).
  • Developing an ABA-inducible split dCas13b for m6A deposition.
  • Utilizing a photoactivatable ABA derivative for light-mediated control.
  • Demonstrating dosage- and time-dependent RNA downregulation.

Main Results:

  • Conditional activation and deactivation of Cas13b/dCas13b activity using ABA.
  • Temporally controlled deposition of m6A at specific RNA sites.
  • Light-induced modulation of split Cas13b/dCas13b system activities.
  • Dosage- and time-dependent downregulation of endogenous RNAs.

Conclusions:

  • The developed split Cas13b/dCas13b systems offer precise temporal and conditional control over RNA manipulation.
  • These platforms expand the CRISPR and RNA regulation toolkit for native cellular environments.
  • The systems minimize functional disruption to endogenous RNAs, facilitating advanced RNA studies.