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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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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 stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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CRISPR Spherical Nucleic Acids.

Chi Huang1, Zhenyu Han1, Michael Evangelopoulos2

  • 1Department of Chemistry and International Institute for Nanotechnology, Northwestern University, Evanston, Illinois 60208-3113, United States.

Journal of the American Chemical Society
|October 6, 2022
PubMed
Summary
This summary is machine-generated.

Researchers developed novel CRISPR Spherical Nucleic Acids (SNAs) for efficient genome editing. These Cas9 ProSNAs enhance cellular delivery and editing efficiency without harsh methods, broadening CRISPR applications.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Nanotechnology

Background:

  • CRISPR/Cas9 genome editing efficiency is hindered by challenges in delivering components into cells and tissues.
  • Spherical nucleic acids (SNAs) offer potential for enhanced cellular uptake but haven't been applied to gene editing.

Purpose of the Study:

  • To design and evaluate a new class of CRISPR SNAs for improved genome editing.
  • To enhance cellular delivery and nuclear localization of CRISPR-Cas9 components.

Main Methods:

  • Synthesized Cas9 ProSNAs: Cas9 protein cores densely modified with DNA exteriors, preloaded with guide RNA.
  • Incorporated GALA peptides and nuclear localization signals for improved endosomal escape and nuclear targeting.
  • Assessed stability against protease digestion and evaluated genome editing efficiency in multiple cell lines.

Main Results:

  • Cas9 ProSNAs demonstrated enhanced cellular uptake without electroporation or transfection agents.
  • The constructs showed stability against protease degradation.
  • Achieved genome editing efficiencies ranging from 32% to 47% across different cell lines.

Conclusions:

  • Novel CRISPR SNAs (Cas9 ProSNAs) overcome key delivery barriers for genome editing.
  • These nanostructures significantly improve cellular uptake and editing efficiency.
  • This advancement holds potential to expand the applications and impact of CRISPR-Cas9 technology.