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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...
131

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Optimized prime editing efficiently generates heritable mutations in maize.

Dexin Qiao1, Junya Wang1, Min-Hui Lu2

  • 1State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.

Journal of Integrative Plant Biology
|December 8, 2022
PubMed
Summary
This summary is machine-generated.

Prime editing efficiency in maize (Zea mays) was significantly improved using the ePE5max system. This breakthrough enables efficient generation of heritable mutations for herbicide resistance traits.

Keywords:
CRISPR/CasMLH1dnepegRNAsherbicide resistancemaizeprime editing

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Area of Science:

  • Plant biotechnology
  • Gene editing technologies
  • Maize genetics

Background:

  • Low efficiency of prime editing (PE) hinders its application in maize.
  • Recent advancements in mammalian cells and rice improved PE efficiency through optimized prime-editing guide RNAs (pegRNAs), prime editor (PE) proteins, and cellular factors.

Purpose of the Study:

  • To evaluate the efficacy of optimized prime editing systems in maize.
  • To overcome the efficiency limitations of prime editing in maize for genetic applications.

Main Methods:

  • Testing of optimized prime editor (PE) systems, including the ePE5max system, in maize.
  • Utilizing PEmax, engineered pegRNAs (pegRNA-evopreQ.1), nicking single guide RNAs (sgRNAs), and MLH1dn.
  • Assessing the generation of heritable mutations conferring herbicide resistance.

Main Results:

  • The ePE5max system demonstrated high efficiency in generating heritable mutations in maize.
  • Successfully created mutations conferring resistance to EPSPS, ALS, and ACCase inhibiting herbicides.
  • Confirmed the system's capability for both homozygous and heterozygous mutations in target genes.

Conclusions:

  • The ePE5max system significantly enhances prime editing efficiency in maize.
  • This advancement removes the primary obstacle to utilizing prime editing in maize.
  • Enables precise genetic modifications for crop improvement and trait development in maize.