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Gene Disruption of Honey Bee Trypanosomatid Parasite, Lotmaria passim, by CRISPR/Cas9 System

Qiushi Liu1, Jing Lei1, Tatsuhiko Kadowaki1

  • 1Department of Biological Sciences, Xi'an Jiaotong-Liverpool University, Suzhou, China.

Frontiers in Cellular and Infection Microbiology
|May 14, 2019
PubMed

Insights

Researchers developed a CRISPR/Cas9 gene-editing method for Lotmaria passim, a parasite infecting honey bees. This new technique allows for targeted gene disruption, aiding the study of honey bee-parasite interactions and improving bee health.

Area of Science:

  • Molecular biology
  • Parasitology
  • Apiculture

Background:

  • Two trypanosomatid species, Lotmaria passim and Crithidia mellificae, infect honey bees.
  • Lotmaria passim is more prevalent and infects the honey bee hindgut.
  • The effects of L. passim infection on honey bee health are not well understood.

Purpose of the Study:

  • To develop a gene-editing method for L. passim using CRISPR/Cas9.
  • To identify genes crucial for L. passim infection and understand their functions.
  • To investigate honey bee-trypanosomatid parasite interactions.

Main Methods:

  • Applied CRISPR/Cas9 system to L. passim.
  • Established L. passim clones expressing tdTomato or Cas9 via electroporation and drug selection.
  • Disrupted miltefosine transporter and tyrosine aminotransferase genes using CRISPR/Cas9-induced homology-directed repair.

Main Results:

  • Successfully created L. passim clones expressing fluorescent markers (tdTomato) or Cas9.
  • Established a method for editing specific L. passim genes.
  • Demonstrated successful gene disruption of miltefosine transporter and tyrosine aminotransferase.

Conclusions:

  • The developed CRISPR/Cas9 system provides a valuable tool for manipulating L. passim.
  • Fluorescently tagged L. passim and gene-editing methods will advance understanding of honey bee-parasite interactions.
  • This research contributes to improving honey bee health by studying parasitic infections.
Keywords:
CRISPR/Cas9Lotmaria passimgenome editinghoney beetrypanosomatid

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