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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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CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art
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Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array.

Bernd Zetsche1,2,3,4,5, Matthias Heidenreich1,2,3,4, Prarthana Mohanraju6

  • 1Broad Institute of MIT and Harvard, Cambridge, MA 02142.

Nature Biotechnology
|December 6, 2016
PubMed
Summary
This summary is machine-generated.

This study simplifies multiplexed genome editing by using Cpf1 (Clustered Regularly Interspaced Short Palindromic Repeats) to process its own CRISPR RNA. This method enables simultaneous editing of multiple genes in mammalian cells and the mouse brain with a single construct.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Multiplexed genome editing with Cas9 is often hindered by the requirement for multiple or large expression constructs.
  • Efficiently targeting several genomic locations simultaneously presents a significant challenge in genetic research and therapeutic development.

Purpose of the Study:

  • To develop a simplified and efficient method for multiplexed genome editing.
  • To leverage the self-processing capability of Cpf1 for streamlined CRISPR array design.

Main Methods:

  • Utilized the Cpf1 nuclease system for its intrinsic CRISPR RNA (crRNA) processing ability.
  • Designed a single, customized CRISPR array for multiplexed gene targeting.
  • Applied the system to edit multiple genes in mammalian cell lines and in vivo in the mouse brain.

Main Results:

  • Demonstrated successful simultaneous editing of up to four genes in mammalian cells using a single Cpf1-based CRISPR array.
  • Achieved simultaneous editing of three genes in the mouse brain, showcasing in vivo applicability.
  • The Cpf1-mediated processing of crRNA significantly simplified the construct design for multiplexed editing.

Conclusions:

  • The Cpf1 system offers a streamlined approach to multiplexed genome editing, overcoming limitations associated with Cas9.
  • This method provides a powerful tool for complex genetic modifications in various biological systems, including in vivo applications.