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Related Concept Videos

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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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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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Developing a CRISPR System in Nongenetic Model Polyploids.

Shengchen Shan1, Bing Yang2,3, Bernard A Hauser4

  • 1Florida Museum of Natural History, University of Florida, Gainesville, FL, USA. shan158538@ufl.edu.

Methods in Molecular Biology (Clifton, N.J.)
|January 31, 2023
PubMed
Summary
This summary is machine-generated.

Whole-genome duplication (WGD) has varied genetic consequences. This study develops CRISPR genome editing for non-model polyploid plants, like Tragopogon, to study gene function after WGD.

Keywords:
CRISPRGenome editingNongenetic modelPlasmid constructionPolyploidyTransformationTransient assay

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Area of Science:

  • Plant genetics
  • Evolutionary biology
  • Genomics

Background:

  • Polyploidy, or whole-genome duplication (WGD), leads to diverse genetic outcomes in organisms.
  • Gene copy loss, divergence, and silencing are common in allopolyploids, impacting their evolution.
  • Functional studies are crucial but challenging in non-model polyploid species due to limited genetic tools.

Purpose of the Study:

  • To adapt and develop CRISPR genome editing tools for non-model polyploid plants.
  • To facilitate functional studies of gene evolution and retention patterns post-WGD.
  • To provide a methodological framework for applying CRISPR in polyploid evolutionary research.

Main Methods:

  • Development of protocols for sgRNA design and plasmid construction for CRISPR.
  • Establishment of a protoplast transient assay for rapid gene function screening.
  • Implementation of a novel plant transformation method for polyploid Tragopogon.
  • Adaptation of protocols for application in other non-model polyploid systems.

Main Results:

  • Demonstrated the feasibility of CRISPR genome editing in diploid and polyploid Tragopogon.
  • Provided detailed protocols for CRISPR implementation in a non-model polyploid system.
  • Showcased the utility of transient assays for gene function analysis in polyploids.
  • Developed a plant transformation method applicable to polyploid Tragopogon.

Conclusions:

  • CRISPR technology can be successfully adapted for functional genomic studies in non-model polyploid plants.
  • The developed protocols offer a valuable guideline for researchers studying WGD in other polyploid species.
  • This work significantly advances the potential for investigating the evolutionary significance of polyploidy using genome editing.