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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 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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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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Updated: Jul 7, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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gRNA Design: How Its Evolution Impacted on CRISPR/Cas9 Systems Refinement.

Cristofer Motoche-Monar1, Julián E Ordoñez1, Oscar Chang2,3

  • 1School of Biological Sciences and Engineering, Yachay Tech University, Urcuquí 100119, Ecuador.

Biomolecules
|December 23, 2023
PubMed
Summary

This study reviews CRISPR/Cas9 guide RNA (gRNA) design tools, tracing their evolution in computational architecture and characteristics. It highlights machine learning and neural networks for optimizing gRNA design in genetic engineering.

Keywords:
CRISPR/Cas9deep learninggRNAmachine learningneural networks

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Area of Science:

  • Molecular Biology
  • Genetics
  • Bioinformatics

Background:

  • The CRISPR/Cas9 system, a bacterial defense mechanism, has been revolutionized into a precise genome editing tool.
  • CRISPR systems offer immunity to bacteria but can be modified for precise genome editing in various organisms.

Purpose of the Study:

  • To provide an overview of historical and recent web-based guide RNA (gRNA) design tools for CRISPR/Cas9.
  • To highlight the evolution of computational architecture and gRNA characteristics in these tools.
  • To emphasize the importance of gRNA design in the accurate performance of CRISPR/Cas9 systems.

Main Methods:

  • Review of existing literature and web-based tools for gRNA design.
  • Analysis of computational approaches, including machine learning and neural networks.
  • Examination of gRNA/target interaction data for prediction and classification.

Main Results:

  • The evolution of gRNA design tools shows advancements in computational architecture and gRNA characteristics.
  • Machine learning and neural networks are increasingly used for predicting and classifying effective gRNAs.
  • Current tools leverage gRNA/target interaction data for improved design accuracy.

Conclusions:

  • Optimized gRNA design is critical for the successful application of CRISPR/Cas9 technology.
  • Advancements in computational approaches and algorithms are essential for developing better gRNAs.
  • This review aims to foster a community focused on utilizing up-to-date algorithms for modern CRISPR/Cas9 engineering.