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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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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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Updated: Feb 17, 2026

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Optimized guide RNA structure for genome editing via Cas9.

Jianyong Xu1,2, Wei Lian1,2, Yuning Jia1,2

  • 1Institute of Biological Therapy, Shenzhen University, Shenzhen, P.R. China.

Oncotarget
|December 8, 2017
PubMed
Summary

Optimizing the guide RNA structure for the Streptococcus pyogenes CRISPR-Cas system enhances genome editing efficiency. A novel chimeric guide RNA with full-length crRNA and tracrRNA sequences demonstrated superior performance.

Keywords:
Cas9RNA guided endonucleasegRNAgenome editingguide RNA

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • The Cas9-guide RNA (gRNA) system is a powerful genome editing tool.
  • Enhancing the efficiency and specificity of CRISPR-Cas genome editing remains a key challenge.

Purpose of the Study:

  • To optimize the guide RNA structure for the Streptococcus pyogenes CRISPR-Cas system.
  • To improve overall genome editing efficiency and specificity.

Main Methods:

  • Investigated different guide RNA structures, including conventional chimeric gRNA and novel forms.
  • Compared the genome editing efficiency of various gRNA structures using the Streptococcus pyogenes CRISPR-Cas system.

Main Results:

  • A chimeric gRNA structure incorporating full-length crRNA and tracrRNA sequences exhibited significantly higher genome editing efficiency.
  • This novel gRNA structure outperformed conventional chimeric gRNAs and other tested variants.

Conclusions:

  • A newly identified gRNA structure with full-length crRNA and tracrRNA enhances Streptococcus pyogenes CRISPR-Cas genome editing efficiency.
  • This finding offers a promising avenue for improving genome editing applications.