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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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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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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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CRISPR/Cas9 Editing of the C. elegans rbm-3.2 Gene using the dpy-10 Co-CRISPR Screening Marker and Assembled Ribonucleoprotein Complexes.
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Base Editing in Peanut Using CRISPR/nCas9.

Anjanasree K Neelakandan1, Binita Subedi2, Sy M Traore2

  • 1Department of Genetics, Development, and Cell Biology, Iowa State University, Ames, IA, United States.

Frontiers in Genome Editing
|June 1, 2022
PubMed
Summary
This summary is machine-generated.

Researchers developed a new base editing tool to increase oleic acid in peanuts. This method efficiently targets key genes, offering a faster way to improve oil quality for health and industry.

Keywords:
CRISPR/nCas9FAD2 genebase editingdeaminasegene-editinghigh oleic acidpoint mutation

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Area of Science:

  • Plant molecular biology and genetics
  • Agricultural biotechnology
  • Food science and nutrition

Background:

  • Peanut (Arachis hypogaea L.) is a vital oilseed crop, with increased oleic acid content desired for health and industrial benefits.
  • The Fatty Acid Desaturase2 (FAD2) genes (AhFAD2A and AhFAD2B) are crucial in regulating oleic acid conversion to linoleic acid.
  • Specific mutations in FAD2 genes can lead to higher oleic acid levels, but traditional breeding methods are time-consuming.

Purpose of the Study:

  • To develop and test novel base editing tools for inducing targeted mutations in peanut AhFAD2 genes.
  • To evaluate the efficiency of two Cas9-deaminase fusion constructs (pDW3873 and pDW3876) for base editing in peanut.

Main Methods:

  • Construction of two expression vectors: pDW3873 (nCas9-PmCDA1) and pDW3876 (rAPOBEC1-UGI-nCas9).
  • Introduction of three guide RNAs (gRNAs) into both constructs to target AhFAD2 genes.
  • Testing the functionality and efficiency of base editing at specific target sites within the AhFAD2 genes.

Main Results:

  • Both pDW3873 and pDW3876 demonstrated base editing activity, successfully replacing cytosine with thymine or other bases.
  • The pDW3873 construct exhibited higher editing efficiency compared to pDW3876.
  • The developed base editing tools effectively targeted both promoter and coding regions of the AhFAD2 genes.

Conclusions:

  • The study presents an efficient base editing system for peanut, with pDW3873 showing superior performance.
  • This tool significantly accelerates the process of introducing beneficial mutations, reducing the typical 15-year introgression timeline.
  • The findings offer substantial benefits for peanut breeders, farmers, the food industry, and consumers by improving oil quality.