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Related Concept Videos

Substrate Generation for Endonucleases of CRISPR/Cas Systems11:53

Substrate Generation for Endonucleases of CRISPR/Cas Systems

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CRISPR/Cas systems mediate adaptive immunity in Bacteria and Archaea. Many Cas proteins are proposed to act as endoribonucleases acting on crRNA precursors of varying length. Here we illustrate three different approaches to generate pre-crRNA substrates for the biochemical analysis of Cas endonuclease...
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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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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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Related Experiment Video

Updated: Jan 20, 2026

Substrate Generation for Endonucleases of CRISPR/Cas Systems
11:53

Substrate Generation for Endonucleases of CRISPR/Cas Systems

Published on: September 8, 2012

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CRISPR-Cas experiments for schools and the public.

Heike Ziegler1, Wolfgang Nellen1

  • 1Science Bridge e.V., Institut für Biologie, Universität Kassel, Heinrich-Plett-Str. 40, D-34132 Kassel, Germany.

Methods (San Diego, Calif.)
|September 1, 2019
PubMed
Summary

Simple CRISPR-Cas gene editing experiments introduce targeted DNA modification to students and non-scientists. These hands-on labs demonstrate CRISPR-Cas9

Keywords:
CRISPRCas9Class room experimentE. coliEducational modulelacZ

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics Education

Background:

  • CRISPR-Cas technology offers revolutionary gene editing capabilities.
  • Broad applications span biomedicine, biotechnology, and agriculture.
  • Public understanding of gene editing is crucial.

Purpose of the Study:

  • To develop simple, accessible CRISPR-Cas experiments for educational purposes.
  • To introduce targeted gene editing to students and non-scientists.
  • To foster a positive attitude towards scientific research.

Main Methods:

  • Part 1: Targeting plasmid-borne lacZ in E. coli to change colony color.
  • Part 2: Analyzing CRISPR-Cas9 induced DNA breaks via colony PCR, gel electrophoresis, or restriction digest.
  • Integrating practical lab work with theoretical background and tutorials.

Main Results:

  • Successful demonstration of targeted gene editing in E. coli.
  • Analysis confirmed CRISPR-Cas9 mediated double-strand breaks in the lacZ gene.
  • The lab course effectively engaged participants aged 16-70.

Conclusions:

  • The developed CRISPR-Cas experiments are robust, inexpensive, and effective for science education.
  • Expert guidance is necessary for successful execution.
  • Full-day lab courses positively influence participants' attitudes toward research.