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Related Concept Videos

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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CRISPR01:59

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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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RNA Interference01:23

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RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
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Related Experiment Video

Updated: Mar 12, 2026

Dissection of Enhancer Function Using Multiplex CRISPR-based Enhancer Interference in Cell Lines
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CRISPR Guide RNA Validation In Vitro.

Stephanie Grainger1,2, Brianna Lonquich1,2, Chet Huan Oon1,2

  • 11 Department of Cellular and Molecular Medicine, University of California , San Diego, La Jolla, California.

Zebrafish
|November 10, 2016
PubMed
Summary
This summary is machine-generated.

We developed an in vitro assay to screen single guide RNA (sgRNA) function for Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 genome editing. This method predicts in vivo editing success, streamlining precise gene modification in zebrafish.

Keywords:
CRISPRCas9in vitroknock invalidationzebrafish

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 technology enables genome editing across diverse model organisms.
  • Precise genomic modifications are often hindered by the challenge of identifying functional single guide RNAs (sgRNAs).

Purpose of the Study:

  • To develop and validate an in vitro method for screening sgRNA efficiency.
  • To improve the reliability and ease of CRISPR/Cas9 genome editing.

Main Methods:

  • An in vitro assay was designed to assess sgRNA function.
  • The assay utilizes common reagents available in zebrafish research laboratories.
  • In vitro results were correlated with in vivo editing outcomes.

Main Results:

  • The developed in vitro assay demonstrated a high correlation with in vivo sgRNA function.
  • The assay successfully predicted the efficacy of sgRNAs for genome editing.
  • The method proved effective across all tested sgRNAs.

Conclusions:

  • This in vitro screening method offers a reliable approach to identify functional sgRNAs for CRISPR/Cas9.
  • The assay simplifies and accelerates the genome editing process.
  • The method, when combined with alternative nucleases and sgRNAs, can facilitate genome editing at nearly any target locus.