C. elegans unc-47 Functional Gene Editing Service

Service Overview Precision editing of unc-47 in a premier genetic model for synaptic transmission, neurotransmitter release and receptor signalling

unc-47 encodes UNC-47, the vesicular GABA transporter. It uses the proton gradient across the synaptic vesicle membrane to pump cytosolic GABA into vesicles, converting transmitter supply into release-ready quanta.

This gene is how VGAT was first identified: the transporter was characterised in the nematode before the vertebrate protein was cloned. In unc-47 mutants exocytosis still occurs but delivers empty vesicles, and cytosolic GABA builds up so immunostaining increases rather than falls. CD BioSciences delivers custom unc-47 alleles — knockouts, transporter point mutants and tag knockins — with full genotype verification.

About the Target Gene: unc-47 Key biological and practical facts to inform your edit design

Gene symbol	unc-47 (the vesicular GABA transporter that pumps cytosolic GABA into synaptic vesicles)
Protein product	UNC-47 — vesicular GABA transporter (VGAT / VIAAT), the vesicular loading step of inhibitory transmission
Mammalian homolog	SLC32A1 (VGAT, vesicular GABA transporter, also called VIAAT)
Biological role	UNC-47 is historically unusual: the vesicular GABA transporter was identified in the worm before it was identified in vertebrates. Molecular analysis showed a protein with ten predicted transmembrane domains, consistent with the transporter superfamilies, produced in GABAergic neurons and absent elsewhere, and mutants accumulated exactly the phenotype predicted for a loading defect. The transporter uses the proton electrochemical gradient generated by the vacuolar ATPase to drive GABA into the vesicle, exchanging protons for transmitter, which is why vesicular filling is coupled to vesicle acidification rather than to ATP directly. This is the step that converts cytosolic GABA into quanta: without it the transmitter is made but never packaged, so a vesicle releases nothing on fusion. Later work showed the same requirement during DD motor neuron remodelling, where both UNC-25 and UNC-47 act to promote timely removal of the ventral synapses laid down in larvae, so GABA signalling feeds back onto its own circuit maturation.
Loss-of-function phenotype	unc-47 mutants are viable and fertile but cannot load GABA into synaptic vesicles. The diagnostic signature runs opposite to intuition: because synthesis continues while packaging fails, GABA accumulates in the cytoplasm and cell bodies actually show elevated rather than reduced GABA immunoreactivity. Functionally the animals behave like GABA-depleted animals, sharing the shrinker phenotype seen in unc-25 mutants, because no GABA reaches the cleft regardless of how much sits in the cytosol. Vesicle fusion itself is normal — exocytosis still happens, it just delivers empty vesicles — which makes this locus the cleanest way to separate the machinery of fusion from the content of release.
Why edit it	VGAT is important far beyond nematodes: it is the reason any inhibitory synapse works, and the human ortholog SLC32A1 is the target of the anti-seizure drug vigabatrin's downstream logic and of experimental approaches to boost inhibition. Editing UNC-47 lets you ask questions merge about quantal content and vesicle filling that cannot be addressed at the synthesis step, and it does so in a viable animal whose entire inhibitory output can be assessed behaviourally in minutes. Because fusion is preserved while content is lost, comparing an unc-47 allele against an unc-25 or snb-1 allele separates transmitter supply, transmitter packaging and release machinery as three experimentally independent variables.
Recommended background	N2 (wild type). Nulls are viable and fertile, so this locus tolerates full deletions. We recommend confirming new alleles by GABA immunostaining, which should show elevated rather than reduced signal, together with a shrinker assay, because that inverted immunostaining result is the most reliable way to distinguish a loading defect from a synthesis defect.
Representative allele	The classical reference alleles from the GABAergic mutant screens plus engineered deletion alleles; custom CRISPR nulls, transporter-domain point mutants that separate substrate recognition from proton coupling, and tag knockins for axonal and vesicle localisation studies.

Fig. 1 UNC-47, the vesicular GABA transporter that pumps cytosolic GABA into synaptic vesicles in C. elegans, the step that makes inhibitory transmission at the neuromuscular junction possible. The highlighted (amber) node marks the editing target.

unc-47 at a glance: unc-47 loss empties vesicles of GABA while leaving exocytosis intact and raising cytosolic GABA levels. Editing unc-47 alone is a powerful way to dissect UNC-47 function in vivo, with conserved relevance to human SLC32A1 biology.

Our unc-47 Editing Strategies Choose the allele that matches your experimental question

Gene Knockout

gRNA-guided Cas9 cutting with NHEJ repair to introduce frameshift indels, generating premature stop codons and null / strong-loss-of-function unc-47 alleles.

Knockin & Tagging

Homology-directed repair (HDR) inserts GFP / HA / FLAG or other tags at the unc-47 locus for endogenous expression and subcellular localization studies.

Point Mutation

Introduce specific amino-acid substitutions to dissect functional domains and separate catalytic from scaffolding roles of UNC-47.

Conditional & Reporter Alleles

Build tissue-specific, inducible, or temperature-sensitive configurations and reporter lines for spatiotemporal analysis of UNC-47 function.

A clean deletion works well because the null is viable and the phenotype is specific, but the most informative design pairs it with an unc-25 allele. If you want to probe transporter chemistry rather than simply remove the protein, point mutants in predicted transmembrane domains let you separate substrate recognition from proton coupling, though these need electrophysiological or immunostaining validation rather than the behavioural assay alone. Always include GABA immunostaining during validation: rising rather than falling signal is the expected result and confirms you have a genuine loading defect.

Workflow of Our unc-47 Editing Service From sequence to validated strain in a streamlined pipeline

Strategy Design

Define target (knockout / knockin / point mutation), select gRNA(s) and HDR template, confirm against the latest genome build.

gRNA Synthesis

Design, validate, and synthesize high-specificity guide RNAs targeting the unc-47 locus.

Vector & Template Construction

Build Cas9 / gRNA delivery construct and, for knockin, the homology-arm repair template.

Microinjection

Perform gonadal microinjection of N2 animals with CRISPR components and co-CRISPR selection markers.

Screening & Verification

PCR, Sanger / sequencing, and phenotype confirmation to isolate edited animals and verify alleles.

Delivery & Report

Provide homozygous / balanced unc-47 strains plus a full genotype and methods report.

Key Features Why researchers choose CD BioSciences for C. elegans editing

High-efficiency, optimized CRISPR / Cas9 platform for C. elegans.
Whole-process technical traceability and transparent timelines.
Flexible editing: knockout, knockin, tagging, and point mutation.
Full genotype verification before strain delivery.
One-stop model development integrated with our C. elegans portfolio.
Competitive pricing and short turnaround.
Direct access to scientists and prompt inquiry response.
Reliable, convenient global shipping of frozen stocks.

Applications Where a custom unc-47 model adds value

Vesicular filling and quantal size.
Proton-coupled neurotransmitter transport.
Separating vesicle fusion from vesicle content.
GABAergic inhibition of body-wall muscle.
The shrinker phenotype and motor coordination.
DD motor neuron synaptic remodelling.
VGAT as a conserved anticonvulsant-relevant target.

Deliverables What you receive with every project

Genetically verified unc-47 edited strain(s) (homozygous or balanced).
Detailed genotyping report (editing strategy, allele sequence, verification data).
Methods summary and rearing / storage recommendations.
Optional: crosses into reporter, glutamatergic / GABAergic / cholinergic neuron marker, and co-CRISPR selection backgrounds.
Optional: phenotype confirmation, aldicarb and levamisole resistance assays, electrophysiological recording at the neuromuscular junction, and locomotor / habituation analysis support.

Frequently Asked Questions

What type of unc-47 allele will I get from a knockout project?

Standard knockout projects deliver a frameshift indel allele predicted to produce a premature stop and a loss-of-function phenotype. We confirm the edit by sequencing and, where relevant, by characteristic phenotype assays.

Can you add a GFP or epitope tag to endogenous unc-47?

Yes. Using HDR with a designed repair template, we can insert GFP, HA, or FLAG tags at the C- or N-terminus while preserving endogenous promoter and regulation.

How long does a typical unc-47 editing project take?

Timelines vary with editing type and validation needs, but our optimized pipeline and co-CRISPR selection typically shorten turnaround versus conventional approaches. Contact us with your design for a project-specific estimate.

Loss of a GABA transporter makes GABA immunostaining go up, not down. Why?

Because the block is in packaging, not production. UNC-25 keeps making GABA from glutamate, but without UNC-47 there is nothing to load it into vesicles, so it accumulates in the cytosol and cell bodies. Staining an unc-47 mutant therefore shows elevated GABA immunoreactivity even though the animal behaves as if GABA signalling were absent, since none of that accumulated transmitter ever reaches the synaptic cleft. unc-25 mutants show the opposite — no detectable GABA at all — because synthesis itself is blocked. That inverted relationship is the fastest way to tell the two loci apart without electrophysiology.

Start Your unc-47 Editing Project

CD BioSciences is committed to convenient, efficient, and cost-effective C. elegans model customization. Tell us your target design — knockout, knockin, tagging, or point mutation — and our scientists will respond with a tailored plan and quote.

References
1. Kim H. M., Colaiácovo M. P. CRISPR-Cas9-guided genome engineering in C. elegans. Current Protocols in Molecular Biology, 2016, 115(1): 31.7.1–31.7.18.
2. McIntire S. L., Reimer R. J., Schuske K., Edwards R. H., Jorgensen E. M. Identification and characterization of the vesicular GABA transporter. Nature, 1997, 389(6653): 870–876.

For research use only.