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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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To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
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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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RNA Editing02:23

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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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CIRCLE-Seq for Interrogation of Off-Target Gene Editing
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[Genome-editing: focus on the off-target effects].

Xiubin He1, Feng Gu1

  • 1State Key Laboratory of Ophthalmology and Optometry, School of Ophthalmology and Optometry, Wenzhou Medical University, Wenzhou 325000, Zhejiang, China.

Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology
|October 31, 2017
PubMed
Summary

Genome editing tools like CRISPR offer therapeutic promise, but unintended DNA alterations, known as off-target effects, remain a significant challenge. This review examines the causes and detection methods for these crucial off-target effects in gene editing.

Keywords:
clustered regulatory interspaced short palindromic repeats (CRISPRs)genome editingoff-targettranscription activator-like effector nucleases (TALENs)zinc-finger nucleases (ZFNs)

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Recent advancements in genome editing technologies, including Zinc-finger nucleases (ZFNs), Transcription activator-like effector nucleases (TALENs), and CRISPR/Cas systems, have opened new avenues for therapeutic strategies.
  • Despite their potential, the occurrence of off-target mutations remains a critical limitation, hindering the safe and effective application of gene editing in clinical settings.

Purpose of the Study:

  • To elucidate the underlying causes of off-target effects in genome editing.
  • To provide a comparative analysis of various methods used for detecting these off-target mutations.

Main Methods:

  • Literature review and synthesis of existing research on genome editing technologies.
  • Comparative analysis of different off-target detection techniques based on their principles, sensitivity, and specificity.

Main Results:

  • Off-target effects arise from the inherent mechanisms of genome editing nucleases recognizing and cleaving DNA sequences similar to their intended targets.
  • Various detection methods, including unbiased sequencing and targeted approaches, exhibit distinct advantages and limitations in identifying off-target mutations.

Conclusions:

  • Understanding the causes of off-target effects is essential for developing strategies to minimize them.
  • Selecting appropriate detection methods is crucial for ensuring the safety and efficacy of genome editing-based therapies.