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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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What is Genetic Engineering?00:49

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Overview
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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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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

Conservative Site-specific Recombination and Phase Variation

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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.
The recognition sites for Cre recombinase called LoxP...
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Related Experiment Video

Updated: Jul 9, 2025

Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

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Recent advances in genome engineering by CRISPR technology.

Youngsik Lee1, Yeounsun Oh1, Seung Hwan Lee1

  • 1Department of Life Science, Chung-Ang University, Seoul 06974, Korea.

BMB Reports
|December 6, 2023
PubMed
Summary
This summary is machine-generated.

CRISPR gene editing is powerful, but unintended off-target edits remain a challenge. This review analyzes CRISPR structures causing these errors and summarizes methods to reduce them.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR technology enables precise gene editing, revolutionizing molecular biology.
  • Off-target mutations, unintended DNA modifications, pose a significant challenge to CRISPR's clinical application.
  • Understanding the structural basis of CRISPR-DNA interactions is crucial for improving specificity.

Approach:

  • This review analyzes the structural principles underlying off-target editing induced by CRISPR systems.
  • It summarizes current methodologies developed to minimize unintended DNA modifications.
  • The focus is on structural aspects influencing CRISPR-Cas9 recognition and activity.

Key Points:

  • CRISPR components inherently possess characteristics that lead to off-target DNA recognition.
  • Structural analysis reveals mechanisms driving unintended cleavage at non-target sites.
  • Various strategies have been developed to enhance CRISPR specificity and reduce off-target effects.

Conclusions:

  • Addressing the off-target problem is essential for the safe and effective future of CRISPR gene editing.
  • Continued research into CRISPR structural biology will guide the development of more precise editing tools.
  • Minimizing off-target edits is critical for advancing CRISPR-based therapeutics and research.