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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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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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Crop cultivation has a long history in human civilization, with records showing the cultivation of cereal plants beginning at around 8000 BC. This early plant breeding was developed primarily to provide a steady supply of food.
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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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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Author Spotlight: Streamlining Rice Breeding with CRISPR/Cas for Obtaining Optimal Phenotypic and Agronomic Traits
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CRISPR/Cas9 Technology for Potato Functional Genomics and Breeding.

Matías Nicolás González1,2, Gabriela Alejandra Massa3,4,5, Mariette Andersson6

  • 1Laboratorio de Agrobiotecnología, IPADS (INTA - CONICET), Balcarce, Argentina. gonzalez.matiasn@inta.gob.ar.

Methods in Molecular Biology (Clifton, N.J.)
|March 30, 2023
PubMed
Summary

CRISPR/Cas9 gene editing offers a powerful method for potato research and breeding. This technology enables precise DNA modifications for functional analysis and improving elite potato cultivars.

Keywords:
Agrobacterium tumefaciensCRISPR/Cas9Crop breedingFunctional genomicsGenome editingPotatoProtoplastsRibonucleoprotein complexes

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Area of Science:

  • Plant Biotechnology
  • Molecular Biology
  • Agricultural Science

Background:

  • Potato (Solanum tuberosum L.) is a vital global staple crop.
  • Its tetraploid and heterozygous nature complicates genetic research and breeding.
  • CRISPR/Cas9 technology offers precise genome editing for potato improvement.

Purpose of the Study:

  • To detail experimental procedures for applying CRISPR/Cas9 in potato genome editing.
  • To provide strategies for target selection and sgRNA design.
  • To facilitate potato gene functional analysis and cultivar improvement.

Main Methods:

  • CRISPR/Cas9 system utilizing single guide RNA (sgRNA) and Cas9 nuclease.
  • Golden Gate-based cloning for sgRNA/Cas9 binary vector construction.
  • Optimized ribonucleoprotein (RNP) complex assembly for protoplast transfection.
  • Agrobacterium-mediated transformation and transient expression in potato protoplasts.
  • Plant regeneration and identification of gene-edited potato lines.

Main Results:

  • Established protocols for CRISPR/Cas9 application in potato.
  • Demonstrated methods for target selection, sgRNA design, and vector construction.
  • Provided optimized RNP complex assembly and transfection protocols.
  • Outlined procedures for identifying gene-edited potato lines.

Conclusions:

  • CRISPR/Cas9 technology is a powerful tool for potato functional genomics.
  • The described methods support efficient gene editing for potato trait improvement.
  • This approach aids in accelerating potato breeding programs.