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Related Concept Videos

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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Related Experiment Video

Updated: Jan 31, 2026

Investigation of the Transcriptional Role of a RUNX1 Intronic Silencer by CRISPR/Cas9 Ribonucleoprotein in Acute Myeloid Leukemia Cells
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Gene Replacement by Intron Targeting with CRISPR-Cas9.

Jun Li1,2, Xiangbing Meng3, Jiayang Li4,5

  • 1State Key Laboratory of Plant Cell and Chromosome Engineering, Center for Genome Editing, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.

Methods in Molecular Biology (Clifton, N.J.)
|January 6, 2019
PubMed
Summary

We developed a new CRISPR-Cas9 method for precise gene replacement in plants. This intron-mediated gene editing technique utilizes non-homologous end joining to efficiently replace targeted DNA fragments.

Keywords:
CRISPR-Cas9DSBGene replacementIntronNHEJ

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Area of Science:

  • Plant biotechnology
  • Genome editing
  • Molecular biology

Background:

  • CRISPR-Cas9 is a powerful genome editing tool, widely used for gene knockouts via non-homologous end joining (NHEJ).
  • Precise gene editing in plants using homology-directed repair (HDR) remains a significant challenge.
  • Targeted gene replacement is crucial for crop improvement and functional genomics.

Purpose of the Study:

  • To develop a novel, efficient method for site-specific gene replacement in plants.
  • To adapt CRISPR-Cas9 technology for precise DNA fragment substitution in plant genomes.
  • To overcome limitations of HDR-based editing in plants.

Main Methods:

  • A new intron-mediated gene replacement strategy was developed using the CRISPR-Cas9 system.
  • Cas9 was engineered to induce simultaneous DNA double-strand breaks (DSBs) in adjacent introns and a donor template.
  • The method leverages the plant's endogenous non-homologous end joining (NHEJ) repair pathway.

Main Results:

  • The developed method enables efficient, site-specific gene fragment replacement in plants.
  • Simultaneous DSB induction in introns and donor DNA facilitates targeted editing.
  • This approach offers a versatile tool for precise genomic modifications in various plant species.

Conclusions:

  • The intron-mediated gene replacement method provides a robust solution for precise plant genome editing.
  • This technique expands the utility of CRISPR-Cas9 for targeted gene modification in plants.
  • The method holds significant potential for advancing plant biotechnology and crop development.