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Related Concept Videos

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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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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CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art
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Multiplexed genome engineering by Cas12a and CRISPR arrays encoded on single transcripts.

Carlo C Campa1, Niels R Weisbach1, António J Santinha1

  • 1Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland.

Nature Methods
|August 14, 2019
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Summary

This study introduces a novel CRISPR-based genome engineering platform enabling simultaneous modification of multiple genes. The system efficiently engineers complex genetic networks for better understanding cellular functions.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Simultaneous genetic modification is crucial for understanding complex cellular functions.
  • Current genome engineering tools have limitations in the scale and type of simultaneous perturbations.

Purpose of the Study:

  • To develop a novel system for simultaneous, multiplexed genome engineering.
  • To overcome the limitations of existing genome engineering technologies.

Main Methods:

  • Encoded Cas12a and a clustered regularly interspaced short palindromic repeat (CRISPR) array in a single transcript using a stabilizer tertiary RNA structure.
  • Utilized up to 25 individual CRISPR RNAs delivered on a single plasmid for multiplexed genome engineering of endogenous targets.

Main Results:

  • Demonstrated successful simultaneous genome engineering using the developed system.
  • Showcased constitutive, conditional, inducible, and orthogonal genome engineering capabilities.
  • Enabled multiplexed engineering of up to 25 targets from a single plasmid.

Conclusions:

  • The developed platform offers a powerful tool for investigating and orchestrating complex genetic programs.
  • This method advances the ability to study gene interactions and networks underlying cellular behaviors.