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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

1.9K
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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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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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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RNA Editing02:23

RNA Editing

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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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DNA as a Genetic Template02:05

DNA as a Genetic Template

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Two structural features of the DNA molecule provide a basis for the mechanisms of heredity: the four nucleotide bases and its double-stranded nature. The Watson-Crick model of double-helical DNA structure, proposed in 1952, drew heavily upon the X-ray crystallography work of researchers Rosalind Franklin and Maurice Wilkins. Watson, Crick, and Wilkins jointly received the Nobel Prize in Physiology or Medicine for their work in 1962. Franklin was, controversially, excluded from the prize for...
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DNA as a Genetic Template02:05

DNA as a Genetic Template

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Related Experiment Video

Updated: Feb 13, 2026

CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation
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CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation

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Easy quantification of template-directed CRISPR/Cas9 editing.

Eva K Brinkman1, Arne N Kousholt2, Tim Harmsen3

  • 1Division of Gene Regulation, Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, the Netherlands.

Nucleic Acids Research
|March 15, 2018
PubMed
Summary
This summary is machine-generated.

Template-directed genome editing with CRISPR/Cas9 introduces specific mutations but requires empirical testing. We developed TIDER, a rapid and accessible method to quantify these editing events, optimizing genome engineering strategies.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Bioengineering

Background:

  • Template-directed CRISPR/Cas9 editing enables precise genomic modifications.
  • Quantifying the efficiency of these edits, especially point mutations, is crucial but challenging.
  • Current methods require empirical validation due to unpredictable success rates.

Purpose of the Study:

  • To adapt the TIDE method for quantifying template-directed CRISPR/Cas9 editing events.
  • To develop a rapid, cost-effective, and accessible tool for optimizing genome editing strategies.
  • To provide a free web tool for analyzing TIDER data.

Main Methods:

  • Adaptation of the Tracking of Insertions/Deletions (TIDE) method.
  • Application to quantify templated editing events, including point mutations.
  • Development of a free web-based analysis tool (http://tide.nki.nl).

Main Results:

  • The TIDER method accurately quantifies templated editing events.
  • It provides a reliable means to assess the success rate of desired mutations.
  • The method is rapid, inexpensive, and accessible for researchers.

Conclusions:

  • TIDER is an effective tool for testing and optimizing template-directed genome editing.
  • This method facilitates the empirical determination of editing efficiency.
  • Accessible tools like TIDER accelerate advancements in genome engineering.