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General Information
Symbol
Hsap\RELAGad1-MI09277-Tp65AD.2
Species
H. sapiens
Name
FlyBase ID
FBal0304271
Feature type
allele
Associated gene
Associated Insertion(s)
Carried in Construct
Also Known As
gad1AD, Gad1MI09277-p65AD
Allele class
Nature of the Allele
Allele class
Mutations Mapped to the Genome
 
Type
Location
Additional Notes
References
Associated Sequence Data
DNA sequence
Protein sequence
 
 
Progenitor genotype
Cytology
Nature of the lesion
Statement
Reference

The attP cassette in the Mi{MIC}Gad1MI09277 insertion has been replaced with a Trojan p65AD cassette that contains a splice acceptor site followed by the T2A peptide, the Hsap\RELA activation domain fused to the T:Zzzz\Zip+ component of the SplitGAL4 system ("p65AD") and an Hsp70 transcription termination signal. The insertion is in the correct orientation to gene trap Gad1. The splice acceptor ensures that the T2A-p65AD open reading frame is incorporated into the Gad1 mRNA, while the T2A sequence is expected to truncate the native Gad1 gene product and promotes the separate translation of the p65AD open reading frame. Thus expression of the Hsap\RELA activation domain fused to the T:Zzzz\Zip+ component of the SplitGAL4 system is driven by the native Gad1 regulatory sequences.

Insertion components
Mi{Trojan-p65AD.2}Gad1MI09277-Tp65AD.2
Product class / Tool use(s)
Expression Data
Reporter Expression
distribution deduced from reporter (Gal4 UAS)
Stage
Tissue/Position (including subcellular localization)
Reference
Additional Information
Statement
Reference

Hsap\RELAGad1-MI09277-Tp65AD.2 in combination with Scer\GAL4dsx-GAL4-DBD drives expression specifically in a subset of GABAergic neurons. These include ~1 adult fruitless pIP-e (male) neuron, ~3 adult doublesex SN (male) neurons and, adult GABAergic doublesex AbN (male) neurons and adult GABAergic doublesex AbN (female) neurons. These latter neurons aborize locally, except for two in males that project to the adult mesothoracic and prothoracic neuromeres. pIP-e neurons and adult doublesex SN (male) neurons project within the superior medial protocerebrum and to the adult subesophageal zone. In males, all pIP-e neurons are positive for fru-XP, as is a single adult doublesex SN (male) neuron and ~30 adult GABAergic doublesex AbN (male) neurons.

 
Marker for
Reflects expression of
Reporter construct used in assay
Human Disease Associations
Disease Ontology (DO) Annotations
Models Based on Experimental Evidence ( 0 )
Disease
Evidence
References
Modifiers Based on Experimental Evidence ( 0 )
Disease
Interaction
References
Comments on Models/Modifiers Based on Experimental Evidence ( 0 )
 
Phenotypic Data
Phenotypic Class
Phenotype Manifest In
Detailed Description
Statement
Reference

Blocking synaptic vesicle exocytosis (using Ctet\TeTxLCTNT.Scer\UAS) in neurons targeted by the split-GAL4 combination of Scer\GAL4dsx-GAL4-DBD and Hsap\RELAGad1-MI09277-Tp65AD.2 does not significantly affect male courtship behavior (they display the normal complement of courtship behaviors including attempts to copulate and relatively normal levels of courtship toward target females), but less than 20% of males successfully copulate (during a 1hr period) and those that do copulate take significantly longer than controls and 80% could not uncouple from the female ('stuck' phenotype); male fertility is also significantly reduced compared to genetic controls. Blocking synaptic vesicle exocytosis (using Ctet\TeTxLCScer\FRT.stop.Scer\UAS) in neurons targeted by the split-GAL4 combination of Scer\GAL4dsx-GAL4-DBD and Hsap\RELAGad1-MI09277-Tp65AD.2 along with Scer\FLP1FLPo.oc.Tag:NLS(SV40-largeT) results in a significant reduction of male courtship behavior but no copulation defects (successful matings in 1hr and copulation duration are not significantly different compared to genetic controls).

Thermogenetic activation (using TrpA1Scer\UAS.(B).cKa at 31[o]C) of neurons targeted by the Split-GAL4 combination of Scer\GAL4dsx-GAL4-DBD and Hsap\RELAGad1-MI09277-Tp65AD.2 does not significantly affect male courtship behavior, but completely blocks genital attachment (no successful matings) in the 1hr observation period. 90% of Scer\GAL4dsx-GAL4-DBD/Hsap\RELAGad1-MI09277-Tp65AD.2 males dismount females and terminate copulation within one minute of thermogenetic activation in copulo; matings truncated in this way never produce progeny. Thermogenetic activation (using TrpA1Scer\FRT.stop.Scer\UAS.T:Hsap\MYC at 31[o]C) of neurons targeted by the Split-GAL4 combination of Scer\GAL4dsx-GAL4-DBD and Hsap\RELAGad1-MI09277-Tp65AD.2 along with Scer\FLP1FLPo.oc.Tag:NLS(SV40-largeT) (both pre-copulation and in copulo) does not significantly impair male courtship (time spent in courtship behavior is similar to genetic controls) or copulation (successful matings in 1hr and copulation duration is similar to genetic controls).

External Data
Interactions
Show genetic interaction network for Enhancers & Suppressors
Phenotypic Class
Phenotype Manifest In
Additional Comments
Genetic Interactions
Statement
Reference
Xenogenetic Interactions
Statement
Reference
Complementation and Rescue Data
Comments
Images (0)
Mutant
Wild-type
Stocks (1)
Notes on Origin
Discoverer
External Crossreferences and Linkouts ( 0 )
Synonyms and Secondary IDs (2)
Reported As
Symbol Synonym
Hsap\RELAGad1-MI09277-Tp65AD.2
Name Synonyms
Secondary FlyBase IDs
    References (6)