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Related Concept Videos

CRISPR01:59

CRISPR

48.8K
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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Updated: May 23, 2025

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins
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Silkworm mutagenesis using a ribonucleoprotein-based CRISPR/Cas12a system.

Yunlong Zou1, Aijun Ye1, Meixin Dong1

  • 1State Key Laboratory of Resource Insects, Key Laboratory of Sericultural Biology and Genetic Breeding, Ministry of Agriculture and Rural Affairs, College of Sericulture, Textile and Biomass Sciences, Southwest University, Chongqing, China.

Insect Biochemistry and Molecular Biology
|May 21, 2025
PubMed
Summary
This summary is machine-generated.

Researchers developed a CRISPR/Cas12a system for silkworm genome editing. This new tool successfully created gene mutations and offers a valuable alternative to existing CRISPR/Cas9 methods for silkworm research.

Keywords:
CRISPR/Cas12aCRISPR/Cas9Cleavage efficiencyGene editingSilkworm

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Area of Science:

  • Genetics
  • Molecular Biology
  • Developmental Biology

Background:

  • CRISPR/Cas9 is a popular genome editing tool in silkworm research.
  • The Cas12a system is a promising alternative but has not been characterized in vivo in silkworms.

Purpose of the Study:

  • To establish and characterize a ribonucleoprotein-based CRISPR/Cas12a system for in vivo genome editing in silkworms.
  • To compare the efficiency of CRISPR/Cas12a with CRISPR/Cas9 in silkworms.
  • To demonstrate the application of CRISPR/Cas12a for generating heritable mutations and observing phenotypes.

Main Methods:

  • Established a ribonucleoprotein-based CRISPR/Cas12a system.
  • Utilized 19 crRNAs and 17 sgRNAs to target three different genes in vivo.
  • Compared Cas12a activity with Cas9.
  • Assessed the effect of temperature (37°C vs. 25°C) on Cas12a activity.
  • Targeted FibH and mp genes to generate mutants.

Main Results:

  • CRISPR/Cas12a generated mutants less efficiently than CRISPR/Cas9.
  • Successfully generated transmissible indels using Cas12a.
  • Produced mutants for FibH and mp genes with expected phenotypes.
  • Demonstrated target-dependent effects of temperature on Cas12a activity.

Conclusions:

  • A functional ribonucleoprotein-based CRISPR/Cas12a system was established in silkworms.
  • CRISPR/Cas12a serves as a practical alternative to CRISPR/Cas9 for silkworm genome editing.
  • This system expands the available genome editing tools for silkworm research.