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Related Concept Videos

CRISPR01:59

CRISPR

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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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Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
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Pooled CRISPR-Based Genetic Screens in Mammalian Cells
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Optimized libraries for CRISPR-Cas9 genetic screens with multiple modalities.

Kendall R Sanson1, Ruth E Hanna1, Mudra Hegde1

  • 1Broad Institute of Harvard and MIT, 75 Ames Street, Cambridge, MA, 02142, USA.

Nature Communications
|December 22, 2018
PubMed
Summary
This summary is machine-generated.

New CRISPR libraries offer superior gene function interrogation. The Brunello (CRISPRko), Dolcetto (CRISPRi), and Calabrese (CRISPRa) tools enhance precision in identifying essential genes and drug resistance mechanisms.

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Area of Science:

  • * Genomics
  • * Molecular Biology
  • * Genetic Engineering

Background:

  • * Genome-wide CRISPR libraries (CRISPR knockout, interference, activation) are crucial for systematic gene function studies.
  • * Existing libraries vary in efficiency for distinguishing essential from non-essential genes.

Purpose of the Study:

  • * To introduce and evaluate novel genome-wide CRISPRko, CRISPRi, and CRISPRa libraries.
  • * To compare their performance against existing tools for gene function interrogation.

Main Methods:

  • * Development and application of Brunello (CRISPRko), Dolcetto (CRISPRi), and Calabrese (CRISPRa) libraries.
  • * Performance assessment using negative and positive selection screens.
  • * Comparison with established CRISPR libraries (GeCKO, SAM) and open reading frame libraries.

Main Results:

  • * Brunello library shows improved performance in distinguishing essential genes compared to previous CRISPRko libraries.
  • * Dolcetto library demonstrates high efficiency in CRISPR interference screens, comparable to CRISPR knockout.
  • * Calabrese library outperforms the SAM approach in CRISPR activation screens for identifying drug resistance genes.

Conclusions:

  • * The developed CRISPR libraries (Brunello, Dolcetto, Calabrese) provide powerful and efficient tools for genome-wide gene function analysis.
  • * These libraries enable multi-modal interrogation of gene function with enhanced precision and performance.