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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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RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
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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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Related Experiment Video

Updated: Aug 15, 2025

Author Spotlight: Streamlining Rice Breeding with CRISPR/Cas for Obtaining Optimal Phenotypic and Agronomic Traits
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Protocol for targeted modification of the rice genome using base editing.

Meixia Wang1, Fang Yan1, Huanbin Zhou2

  • 1State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.

STAR Protocols
|January 3, 2023
PubMed
Summary

This study details a protocol for precise adenine base editing (ABE) in rice, enabling targeted adenine-to-guanine substitutions. This method facilitates the generation of single-nucleotide variants for genetic research.

Keywords:
Biotechnology and bioengineeringCRISPRGeneticsGenomicsMolecular BiologyPlant sciencesSequencing

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Area of Science:

  • Plant Biotechnology
  • Genome Editing
  • Molecular Biology

Background:

  • Base editing offers precise genome modification for generating single-nucleotide variants (SNVs).
  • Adenine Base Editors (ABE) enable targeted A-to-G substitutions, a crucial tool in genetic engineering.
  • Efficient genome editing protocols are essential for advancing crop improvement in rice.

Purpose of the Study:

  • To present a detailed protocol for adenine base editing (ABE) in rice.
  • To enable targeted adenine (A)-to-guanine (G) substitutions in the rice genome.
  • To provide a foundation for cytosine base editing applications in rice.

Main Methods:

  • Design of single-guide RNA (sgRNA) for target specificity.
  • CRISPR plasmid construction for ABE delivery.
  • Rapid genetic transformation and efficient genotyping of editing events in rice.

Main Results:

  • Successful implementation of a protocol for A-to-G base editing in rice using ABE.
  • Demonstration of key steps including sgRNA design, plasmid construction, and transformation.
  • Validation of the protocol's applicability for generating SNVs in rice.

Conclusions:

  • The presented protocol enables precise A-to-G base editing in rice.
  • This methodology is adaptable for cytosine base editing in rice.
  • The protocol facilitates the generation of targeted genetic modifications in rice for research and breeding.