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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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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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Updated: Oct 2, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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SgRNA engineering for improved genome editing and expanded functional assays.

Chang Dong1, Yuanwei Gou1, Jiazhang Lian1

  • 1Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China; Hangzhou Global Scientific and Technological Innovation Center, Zhejiang University, Hangzhou 310027, China.

Current Opinion in Biotechnology
|February 26, 2022
PubMed
Summary
This summary is machine-generated.

Engineered single guide RNAs (sgRNAs) enhance CRISPR/Cas genome editing activity and specificity. Modifications enable new CRISPR-based applications like transcriptional regulation and prime editing for advanced biotechnology.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genome Engineering

Background:

  • The CRISPR/Cas system is a leading tool for genome engineering in research and biotech.
  • Significant research focuses on improving CRISPR system performance and functions.
  • Single guide RNAs (sgRNAs) are crucial components, with ongoing efforts in their design and engineering.

Purpose of the Study:

  • To review advancements in engineering sgRNA nucleotide sequences for enhanced CRISPR/Cas genome editing.
  • To highlight how sgRNA modifications expand CRISPR-based functional assays.
  • To discuss future directions in sgRNA engineering for more potent CRISPR systems.

Main Methods:

  • Review of literature on sgRNA sequence optimization for improved activity and specificity.
  • Analysis of studies incorporating aptamers and end extensions into sgRNAs.
  • Examination of applications such as transcriptional regulation, genome imaging, and prime editing.

Main Results:

  • Optimized sgRNA sequences demonstrably improve genome editing efficiency and precision.
  • Incorporation of aptamers and extensions enables novel CRISPR-based functional applications.
  • Engineered sgRNAs expand the versatility of CRISPR/Cas systems beyond basic gene editing.

Conclusions:

  • sgRNA engineering is key to advancing CRISPR/Cas technology.
  • Further innovations in sgRNA design will unlock more powerful and versatile genome engineering tools.
  • The field is moving towards sophisticated CRISPR applications through sgRNA modifications.