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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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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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Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Efficient Genome Editing in Multiple Salmonid Cell Lines Using Ribonucleoprotein Complexes.

Remi L Gratacap1, Ye Hwa Jin1, Marina Mantsopoulou1

  • 1The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Midlothian, EH25 9RG, UK.

Marine Biotechnology (New York, N.Y.)
|September 18, 2020
PubMed
Summary
This summary is machine-generated.

Genome editing in salmonid cell lines is now possible using ribonucleoprotein complexes. This breakthrough facilitates genetic studies for disease resistance in aquaculture, benefiting Atlantic salmon and rainbow trout.

Keywords:
CRISPRCell lineDisease resistanceGenome editingRibonucleoproteinSalmonid

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Area of Science:

  • Aquaculture
  • Genomics
  • Molecular Biology

Background:

  • Infectious diseases significantly impact salmonid aquaculture economics and animal welfare.
  • Understanding host response and genetic resistance is crucial for disease prevention and treatment.
  • Cell lines are valuable models for studying salmonid infectious diseases, but efficient genome editing methods are lacking for key species.

Purpose of the Study:

  • To optimize and test a genome editing method using ribonucleoprotein (RNP) complexes in commonly used salmonid cell lines.
  • To establish efficient gene editing protocols for Atlantic salmon and rainbow trout cell lines, overcoming previous transduction difficulties.

Main Methods:

  • Utilized ribonucleoprotein (RNP) complexes for genome editing.
  • Employed electroporation for delivering RNPs into cell lines.
  • Tested both Cas9 and Cas12a enzymes for editing efficiency.
  • Applied the method to Atlantic salmon (SHK-1, ASK), rainbow trout (RTG-2), and Chinook salmon (CHSE-214) cell lines.

Main Results:

  • Achieved efficient and targeted genome editing in all tested salmonid cell lines, with typically over 90% of cells edited.
  • Demonstrated the effectiveness of RNP-based genome editing via electroporation.
  • Showcased the utility of both Cas9 and Cas12a, expanding potential editing targets.

Conclusions:

  • Optimized RNP-based genome editing protocols are now available for key salmonid cell lines.
  • These protocols will significantly advance functional genetic studies in salmonid aquaculture.
  • Enables enhanced research into disease resistance mechanisms in commercially important fish species.