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Can genetic engineering-based methods for gene function identification be eclipsed by genome editing in plants? A

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Comparing plant gene function identification methods is crucial for crop improvement. This review analyzes eight techniques, including CRISPR/Cas9, highlighting their pros and cons for effective gene discovery.

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Area of Science:

  • Plant biology
  • Genetics
  • Molecular biology

Background:

  • Identifying plant gene functions is vital for crop improvement and bioprospecting.
  • Numerous techniques exist, with clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9) being a popular genome editing method.
  • However, CRISPR/Cas9 has limitations, necessitating a comparison with other established and emerging technologies.

Purpose of the Study:

  • To comprehensively compare eight distinct plant gene function identification technologies.
  • To analyze the advantages, disadvantages, similarities, and dissimilarities of these methods across 14 parameters.
  • To provide a guide for researchers in selecting appropriate techniques for crop improvement and bioprospecting.

Main Methods:

  • Review and comparison of eight gene function identification technologies in plants.
  • Categorization of methods into genetic engineering, transient expression, and chemical/physical mutagenesis.
  • Evaluation based on 14 distinct parameters, including efficacy and limitations.

Main Results:

  • CRISPR/Cas9 excels at gene sequence modification and allelic variation but can cause off-target mutations.
  • Other methods offer targeted gene knockout, knockdown, or overexpression, addressing limitations of genome editing.
  • Strategies to overcome shortcomings of targeted gene knockout and CRISPR/Cas9 methods are discussed.

Conclusions:

  • No single method is universally optimal for plant gene function identification.
  • A thorough understanding of each technology's strengths and weaknesses is essential for successful application.
  • This comparative review aids researchers in making informed decisions for plant science research and applications.