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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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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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Homologous Recombination02:31

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Conservative Site-specific Recombination and Phase Variation02:53

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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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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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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
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Targeted genomic rearrangements using CRISPR/Cas technology.

Peter S Choi1, Matthew Meyerson2

  • 11] Dana-Farber Cancer Institute, Department of Medical Oncology, Boston, Massachusetts 02215, USA [2] Broad Institute of MIT and Harvard, Cancer Program, Cambridge, Massachusetts 02142, USA.

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|April 25, 2014
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Summary
This summary is machine-generated.

CRISPR/Cas technology enables precise generation of cancer-related chromosomal rearrangements, like CD74-ROS1 translocations and EML4-ALK inversions. This breakthrough facilitates functional analysis of genomic rearrangements in lung cancer research.

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Area of Science:

  • Genetics and Genomics
  • Cancer Biology
  • Molecular Biology

Background:

  • Genomic rearrangements are common in cancer but challenging to engineer for study.
  • Specific chromosomal alterations drive cancer development, particularly in lung cancer.
  • Functional analysis of these rearrangements is crucial for understanding oncogenesis.

Purpose of the Study:

  • To demonstrate the utility of CRISPR/Cas technology for generating specific chromosomal rearrangements.
  • To create models for studying driver events in lung cancer.
  • To provide a tractable method for investigating genomic rearrangements.

Main Methods:

  • Utilized CRISPR/Cas9 gene editing technology.
  • Targeted specific loci to induce DNA breaks.
  • Induced chromosomal translocations and inversions implicated in lung cancer.

Main Results:

  • Successfully generated CD74-ROS1 translocations.
  • Successfully generated EML4-ALK and KIF5B-RET inversions.
  • Demonstrated efficient rearrangement between targeted loci via Cas9-induced breaks.

Conclusions:

  • CRISPR/Cas technology offers a precise method for creating complex genomic rearrangements.
  • This approach enables functional studies of cancer-driving chromosomal alterations.
  • The method provides a valuable tool for cancer genomics research.