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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

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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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Genome Editing in the Yellow Fever Mosquito Aedes aegypti using CRISPR-Cas9
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Programmable CRISPR interference for gene silencing using Cas13a in mosquitoes.

Aditi Kulkarni1, Wanqin Yu1, Alex S Moon1

  • 1Department of Biology, New Mexico State University, PO Box 30001 MSC 3AF, Las Cruces NM, 88003, USA.

Journal of Genomics
|March 20, 2020
PubMed
Summary

Researchers developed a CRISPR interference tool using Cas13a to silence specific genes in mosquitoes, offering a new method for gene knockdown with no observed off-target effects.

Keywords:
Aedes aegyptiAnopheles gambiaeCRISPR-Cas13aCRISPRiRNA interferencegene silencing

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Area of Science:

  • Molecular Biology
  • Genetics
  • Vector Control

Background:

  • CRISPR-Cas systems offer precise gene editing capabilities.
  • Cas13a is an RNA-guided RNA nuclease targeting single-stranded RNA.
  • Gene silencing in mosquitoes is crucial for vector control research.

Purpose of the Study:

  • To develop and evaluate a Cas13a-mediated CRISPR interference (CRISPRi) tool for gene silencing in mosquitoes.
  • To assess the efficacy and specificity of this tool in different mosquito species.

Main Methods:

  • Development of a Cas13a-expressing plasmid for intrathoracic injection into mosquitoes.
  • Synthesis of target-specific CRISPR RNA (crRNA) in vitro.
  • Delivery of Cas13a and crRNA to Anopheles gambiae and Aedes aegypti.
  • Analysis of gene silencing effects on target genes (vitellogenin, COPI subunits) and organismal phenotypes.

Main Results:

  • Cas13a transcripts were detected for at least 10 days post-injection.
  • Successful silencing of the vitellogenin gene in Anopheles gambiae led to reduced egg production.
  • Silencing of COPI gene subunits in Aedes aegypti resulted in mortality and midgut fragility.
  • Simultaneous gene silencing was achieved using a crRNA cocktail.
  • No significant collateral cleavage of non-target transcripts was observed.

Conclusions:

  • Cas13a-mediated CRISPR interference is a viable and programmable tool for gene knockdown in mosquitoes.
  • This method provides an alternative to traditional RNA interference (RNAi) techniques like dsRNA or siRNA.
  • The CRISPRi system demonstrates high specificity and potential for applications in mosquito research and control.