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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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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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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Two efficient CRISPR/Cas9 systems for gene editing in soybean.

Jéssica Carrijo1,2, Eudald Illa-Berenguer3, Peter LaFayette3,4

  • 1Embrapa Recursos Genéticos e Biotecnologia, Parque Estação Biológica, PqEB, Av W5 Norte Final 716, Brasília, DF, 70770-917, Brazil.

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|April 2, 2021
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Summary

This study compares two CRISPR/Cas9 systems for soybean genome editing. The two-component system (TCTU) showed higher efficiency for editing GmIPK1 and GmIPK2 genes than the single transcriptional unit (STU) system.

Keywords:
Genome editingLow phytic acidSingle transcriptional unitTwo-component transcriptional unit

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Area of Science:

  • Agricultural Science
  • Molecular Biology
  • Biotechnology

Background:

  • CRISPR/Cas9 is a key tool for crop improvement, but efficiency varies in complex genomes like soybean.
  • Optimizing CRISPR/Cas9 delivery and expression is crucial for enhancing gene editing outcomes in plants.

Purpose of the Study:

  • To compare the efficiency of a single transcriptional unit (STU) versus a two-component transcriptional unit (TCTU) CRISPR/Cas9 system for soybean genome editing.
  • To evaluate these systems for multiplex gene editing targeting GmIPK1 and GmIPK2 involved in phytic acid synthesis.

Main Methods:

  • Utilized CRISPR/Cas9 technology with two distinct systems: STU (SpCas9 and sgRNA from one promoter) and TCTU (SpCas9 and sgRNA from separate promoters).
  • Employed a multiplex system targeting two soybean genes (GmIPK1, GmIPK2) using the hairy root transformation method.

Main Results:

  • Both STU and TCTU systems achieved gene-specific editing in soybean.
  • The TCTU system demonstrated higher indel frequencies for GmIPK1 editing (1-749 bp deletions) compared to STU.
  • While both systems showed major exclusions for GmIPK2, STU exhibited lower editing efficiency.

Conclusions:

  • Both STU and TCTU CRISPR/Cas9 systems are effective for soybean gene editing.
  • The TCTU system is more efficient overall, whereas the STU system offers a smaller CRISPR/Cas cassette size.