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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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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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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.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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Related Experiment Video

Updated: May 17, 2025

CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation
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CRISPR/Cas9 Delivery Systems to Enhance Gene Editing Efficiency.

Ana Seijas1,2, Diego Cora2, Mercedes Novo2

  • 1Departamento de Zooloxía, Xenética e Antropoloxía Física, Facultade de Veterinaria, Universidade de Santiago de Compostela, 27002 Lugo, Spain.

International Journal of Molecular Sciences
|May 14, 2025
PubMed
Summary

This review explores CRISPR/Cas9 gene editing delivery systems, focusing on physical, viral, and non-viral methods. It addresses challenges like off-target effects and protein aggregation to advance safer CRISPR/Cas9 therapies.

Keywords:
CRISPRCas9Cas9 aggregationLNPsdelivery systemsgene editingnanoparticle encapsulationnon-viral vectorsphysical delivery

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Gene Therapy

Background:

  • CRISPR/Cas9 technology offers precise genome editing with significant therapeutic promise.
  • Key challenges include off-target effects, immunogenicity, and efficient delivery of CRISPR components.
  • Cas9 protein aggregation can hinder cellular uptake, encapsulation, and nuclear localization, impacting therapeutic efficacy.

Purpose of the Study:

  • To provide an integrated analysis of CRISPR/Cas9 delivery systems.
  • To highlight advances in physical, viral, and non-viral delivery platforms.
  • To identify critical parameters for successful CRISPR/Cas9-based therapies and propose standardized assessment methods.

Main Methods:

  • Review of physical, viral, and non-viral delivery systems for CRISPR/Cas9 components.
  • Analysis of recent advances in lipid nanoparticles, polymeric carriers, and hybrid platforms.
  • Examination of Cas9 protein aggregation and its impact on delivery and editing outcomes.

Main Results:

  • Comparison of delivery platforms and their editing outcomes reveals critical physicochemical parameters.
  • Identification of Cas9 aggregation as a significant factor affecting delivery efficiency and nuclear localization.
  • Insights into challenges related to manufacturing scalability and regulatory requirements for CRISPR/Cas9 therapies.

Conclusions:

  • Optimizing delivery systems and addressing Cas9 aggregation are crucial for effective CRISPR/Cas9 therapies.
  • Standardized methods for assessing Cas9 encapsulation and aggregation are needed.
  • Overcoming translational barriers is essential for the clinical application of CRISPR/Cas9 technology.