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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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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 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: Oct 1, 2025

Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids
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Antispacer peptide nucleic acids for sequence-specific CRISPR-Cas9 modulation.

Nicholas G Economos1,2, Elias Quijano1,2, Kelly E W Carufe1,2

  • 1Department of Therapeutic Radiology, Yale School of Medicine, New Haven, CT 06520 USA.

Nucleic Acids Research
|March 2, 2022
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Summary
This summary is machine-generated.

New antispacer peptide nucleic acids (PNAs) offer precise control over CRISPR-Cas9 gene editing. These tools enable sequence-specific modulation of Cas9 activity, enhancing versatility and safety in various applications.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Gene Editing Technologies

Background:

  • CRISPR-Cas9 technology is widely used but lacks tools for precise dose, timing, and specificity control.
  • Synthetic peptide nucleic acids (PNAs) bind RNA with high affinity, offering a basis for novel molecular tools.

Purpose of the Study:

  • To develop and characterize antispacer PNAs for modulating CRISPR-Cas9 binding and activity.
  • To demonstrate sequence-specific inhibition and regulation of Cas9 and dCas9-fusion systems.

Main Methods:

  • Design and synthesis of guide RNA (gRNA) spacer-targeted antispacer PNAs.
  • Testing PNA efficacy in inhibiting Cas9 binding and activity in cellular systems.
  • Application of antispacer PNAs for temporal control of dCas9-fusion proteins.

Main Results:

  • Antispacer PNAs efficiently target gRNA spacer sequences at low doses, inhibiting Cas9 activity sequence-selectively.
  • Short PAM-proximal PNAs achieved over 2000-fold cleavage inhibition.
  • PAM-distal PNAs modulated gRNA affinity, enhancing on-target specificity.
  • Antispacer PNAs enabled temporal regulation of dCas9-fusion systems.

Conclusions:

  • Antispacer PNAs represent a novel approach to nucleoprotein engineering.
  • This platform offers rapid implementation for CRISPR-Cas9 modulation, improving spatiotemporal control and safety.