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Related Concept Videos

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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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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Related Experiment Video

Updated: Jul 24, 2025

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
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Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

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Simultaneous multifunctional transcriptome engineering by CRISPR RNA scaffold.

Zukai Liu1,2, Nathaniel Jillette1, Paul Robson1,2,3,4

  • 1The Jackson Laboratory for Genomic Medicine, Farmington, CT 06032, USA.

Nucleic Acids Research
|July 3, 2023
PubMed
Summary
This summary is machine-generated.

A new platform called Combinatorial RNA Engineering via Scaffold Tagged gRNA (CREST) enables simultaneous manipulation of multiple RNA targets. This system significantly reduces off-target effects, enhancing RNA engineering capabilities.

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Area of Science:

  • Molecular Biology
  • RNA Biology
  • Biotechnology

Background:

  • Precise regulation of RNA processing and metabolism is crucial for cellular integrity and function.
  • CRISPR-Cas13 systems allow targeted RNA engineering, but simultaneous modulation of multiple RNA processing steps and off-target effects remain challenges.

Purpose of the Study:

  • To develop a novel platform for simultaneous, multi-functional RNA modulation.
  • To overcome limitations of existing RNA engineering tools, including off-target events.

Main Methods:

  • Developed Combinatorial RNA Engineering via Scaffold Tagged gRNA (CREST) platform.
  • Appended RNA scaffolds to Cas13 gRNA and fused cognate RNA binding proteins with enzymatic domains.
  • Created bifunctional and tri-functional CREST systems for RNA alternative splicing and base editing (A-to-G, C-to-U).
  • Utilized a split-design approach for enzyme reconstitution to minimize off-target events.

Main Results:

  • Demonstrated simultaneous RNA manipulation of alternative splicing and base editing.
  • Achieved significant reduction (nearly 99%) in off-target events using a split-enzyme design.
  • Successfully reconstituted enzyme activity at target sites.

Conclusions:

  • CREST platform enables simultaneous execution of multiple RNA modulation functions on different RNA targets.
  • The split-design strategy effectively minimizes off-target effects, improving specificity.
  • CREST expands the transcriptome engineering toolbox for RNA biology research.