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Hox gene-specific cellular targeting using split intein Trojan exons.

Fengqiu Diao1, Deeptha Vasudevan2, Ellie S Heckscher2

  • 1Laboratory of Molecular Biology, Section on Neural Function, National Institute of Mental Health, NIH, Bethesda, MD 20892.

Proceedings of the National Academy of Sciences of the United States of America
|April 11, 2024
PubMed
Summary
This summary is machine-generated.

A new Split Intein Trojan exon (siTrojan) method improves Drosophila Gal4 driver lines by preventing harmful mutations. This technique ensures full-length protein production, enabling precise gene targeting for cell type analysis.

Keywords:
Drosophilacell typedevelopmentgenetic accessneural circuit-mapping

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Area of Science:

  • Developmental Biology
  • Genetics
  • Neuroscience

Background:

  • The Trojan exon method in Drosophila generates gene-specific Gal4 driver lines for cell type access and loss-of-function analysis.
  • However, truncation mutations from Trojan exons can cause dominant-negative effects and developmental lethality, especially for essential transcription factors.
  • Alternative methods are needed to create Gal4 lines targeting transcription factors without detrimental mutations.

Purpose of the Study:

  • To introduce a modified Trojan exon method that mitigates mutagenic effects while retaining targeting specificity.
  • To develop a toolkit of Gal4 and Split Gal4 lines for Hox transcription factors.
  • To demonstrate the utility of the new method in neural circuit mapping.

Main Methods:

  • Developed Split Intein Trojan exons (siTrojans) that utilize self-splicing split inteins.
  • siTrojans ensure trans-splicing of truncation products to restore full-length native protein function.
  • Generated Gal4 and Split Gal4 lines for segmentally expressed Hox transcription factors.

Main Results:

  • The siTrojan method successfully mitigates mutagenic effects of the original Trojan exon approach.
  • A comprehensive toolkit of Gal4 and Split Gal4 lines for Hox transcription factors was created.
  • Demonstrated application of siTrojans in neural circuit mapping by targeting neurons based on anterior-posterior position.

Conclusions:

  • Split Intein Trojan exons (siTrojans) offer a valuable alternative for generating gene-specific Gal4 driver lines in Drosophila.
  • This method preserves targeting fidelity and modularity while preventing deleterious mutations.
  • The siTrojan method and the Hox transcription factor toolkit provide broad utility for genetic studies in Drosophila.