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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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CRISPR/Cas9 Genome Editing01:28

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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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What is Genetic Engineering?00:49

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CRISPR and crRNAs02:53

CRISPR and crRNAs

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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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Homologous Recombination02:31

Homologous Recombination

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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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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
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Engineered CRISPR Systems for Next Generation Gene Therapies.

Michael Pineda1, Farzaneh Moghadam1, Mo R Ebrahimkhani1,2

  • 1School of Biological and Health Systems Engineering, Arizona State University , Tempe, Arizona 85281, United States.

ACS Synthetic Biology
|May 31, 2017
PubMed
Summary

CRISPR gene editing offers precise in vivo therapy but faces challenges in control, delivery, and efficiency. Bioengineering advancements are crucial for translating this powerful genome engineering tool into clinical applications.

Keywords:
CRISPRCas9clustered regularly interspaced short palindromic repeatscontrollable CRISPRgene editinggene modulation,gRNAgene therapysafe CRISPRsynthetic biology

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics

Background:

  • Gene therapy aims for safe, precise in vivo modulation of gene regulatory networks.
  • Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) enable targeted DNA modification.
  • CRISPR-Cas9 systems offer flexibility and precision through guide-RNA (gRNA) complementarity to genomic sequences.

Purpose of the Study:

  • To evaluate CRISPR's potential as a next-generation in vivo gene therapy platform.
  • To discuss bioengineering advancements addressing challenges in CRISPR clinical translation.

Main Methods:

  • Review of CRISPR-Cas9 system components (Cas9 protein, gRNA, delivery platforms).
  • Analysis of engineered modifications to CRISPR system components (DNA, RNA, ribonucleoprotein).

Main Results:

  • CRISPR-Cas9 holds significant promise for in vivo genetic engineering and therapeutics.
  • Key challenges include minimizing off-target effects, maximizing editing efficiency, and achieving spatial-temporal regulation.
  • Independent engineering of CRISPR components has led to improved genome engineering tools.

Conclusions:

  • CRISPR-Cas9 is a powerful tool with vast applications in genetic disease modeling and synthetic gene regulation.
  • Overcoming current hurdles is essential for the clinical implementation of CRISPR-based in vivo gene therapies.
  • Continued bioengineering efforts are vital for advancing CRISPR technology towards safe and effective therapeutic use.