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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

746
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

CRISPR

53.7K
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/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art
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Multiplexed Genome Engineering with Cas12a.

Niels R Weisbach1, Ab Meijs1, Randall J Platt2,3,4

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

Methods in Molecular Biology (Clifton, N.J.)
|July 6, 2021
PubMed
Summary
This summary is machine-generated.

CRISPR-Cas12a technology enables simultaneous engineering of hundreds of genes for studying complex cellular programs. This massively multiplexed genome engineering approach offers a novel way to interrogate and engineer genetic functions in eukaryotic cells.

Keywords:
CRISPRCRISPR array synthesisCRISPR-Cas12aCas12aGene editingGenome engineeringMultiplexedOrthogonal genome engineeringTranscriptional regulation

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR-Cas systems facilitate high-throughput study of genotype-phenotype relationships.
  • Existing genome engineering tools are limited to manipulating one or a few genes at a time.
  • Engineering complex cellular programs requires integrated genome manipulation technologies.

Purpose of the Study:

  • To introduce single transcript CRISPR-Cas12a (SiT-Cas12a) for massively multiplexed genome engineering.
  • To enable conditional, inducible, and orthogonal manipulation of numerous genomic targets simultaneously.
  • To provide a unified framework for engineering complex genetic programs in eukaryotic cells.

Main Methods:

  • Development and application of SiT-Cas12a in human cells.
  • Design and assembly of customized multiplexed CRISPR guide RNA arrays.
  • Validation and analysis of guide RNA array processing and genome engineering outcomes.

Main Results:

  • SiT-Cas12a enables massively multiplexed genome engineering of dozens to hundreds of targets.
  • Demonstrated utility of SiT-Cas12a for interrogating complex genetic programs.
  • Established experimental procedures for SiT-Cas12a application.

Conclusions:

  • SiT-Cas12a is a powerful tool for simultaneous, large-scale genome engineering.
  • This technology advances the study and engineering of complex cellular functions.
  • Provides a novel approach for high-throughput genetic analysis and manipulation.