C. elegans unc-54 Functional Gene Editing Service

Service Overview Precision editing of unc-54 in a premier genetic model for muscle contractility, neuronal wiring, and locomotion

unc-54 encodes myosin heavy chain B (MHC B) of Caenorhabditis elegans, the principal motor of the body-wall muscle and a member of the sarcomeric myosin heavy chain family whose human counterparts include MYH1–MYH8.

Within a single thick filament, MHC A occupies the central zone and MHC B the polar regions, so the two heavy chains build one structure rather than two. Historically this locus is where myosin was first tied to muscle function by mutation, and it remains a standard platform for dissecting motor activity, filament assembly and myopathy variants. CD BioSciences delivers custom unc-54 alleles — knockouts, ATPase-defective motors, rod truncations, human MYH variant knockins, and tag knockins — with full genotype verification.

About the Target Gene: unc-54 Key biological and practical facts to inform your edit design

Gene symbol	unc-54 (Myosin heavy chain B (MHC B), the major body-wall muscle motor)
Protein product	UNC-54 — myosin heavy chain B of the body-wall muscle
Mammalian homolog	Skeletal muscle myosin heavy chain family (MYH1, MYH2, MYH3, MYH4, MYH7, MYH8 class)
Biological role	UNC-54 is the motor that does most of the work in C. elegans body-wall muscle. Each sarcomere contains a single class of thick filament, and within that filament MHC A (MYO-3) occupies the central zone while MHC B (UNC-54) fills the two polar regions, so the two heavy chains are not interchangeable but occupy complementary positions along the same bipolar filament. Each heavy chain carries the N-terminal motor head that hydrolyses ATP and walks along actin, plus the long coiled-coil rod that assembles the filament backbone.
Loss-of-function phenotype	Removing UNC-54 removes most of the contractile machinery of the body wall. Animals are strongly paralysed and cannot sustain sinusoidal movement; the thick filaments that remain are dominated by MHC A and fail to build normal, full-length assemblies. Because the defect is structural rather than lethal, you can still recover adults and assay their muscle directly.
Why edit it	This is the gene in which myosin was first connected to muscle function by mutation, and it remains one of the most tractable myosin systems anywhere: the two heavy chains are genetically separable but physically co-assembled, so you can dissect motor activity and filament assembly independently in the same cell. Human MYH mutations cause a spectrum of skeletal myopathies and cardiomyopathies, which makes a whole-animal myosin model directly relevant.
Recommended background	N2 (wild type). Null animals are recoverable; we recommend scoring swimming and crawling rates in parallel with sarcomere organisation, since partial loss-of-function and null alleles can diverge. If you plan any work on filament assembly, combine the edit with a MYO-3 or paramyosin reporter.
Representative allele	Classic reference alleles including the widely used missense allele e1092 and null alleles, plus engineered deletions; custom CRISPR nulls, ATPase-defective motor mutants, rod truncations, human-variant knockins, and fluorescent or epitope tag knockins available on request.

Fig. 1 UNC-54 / myosin heavy chain B, the major motor that powers contraction of the C. elegans body-wall muscle. The highlighted (amber) node marks the editing target.

unc-54 at a glance: unc-54 supplies most of the body-wall myosin motor, so nulls are paralysed and barely able to move. Editing unc-54 alone is a powerful way to dissect UNC-54 function in vivo, with conserved relevance to human Skeletal muscle myosin heavy chain family biology.

Our unc-54 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-54 alleles.

Knockin & Tagging

Homology-directed repair (HDR) inserts GFP / HA / FLAG or other tags at the unc-54 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-54.

Conditional & Reporter Alleles

Build tissue-specific, inducible, or temperature-sensitive configurations and reporter lines for spatiotemporal analysis of UNC-54 function.

An ATPase-defective motor mutant is usually more informative than a deletion: the protein is still built into the filament, so you separate catalysis from assembly. One caveat worth planning around — because MHC A remains, single nulls can retain residual filament structures, so if you want the clean assembly defect, consider combining this edit with a myo-3 lesion.

Workflow of Our unc-54 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-54 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-54 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-54 model adds value

Myosin motor activity and ATPase function in vivo.
Thick filament assembly and bipolar filament length control.
Division of labour between MHC A and MHC B within one filament.
Sliding filament mechanics and force generation.
Human sarcomeric myosin variant modelling in a whole animal.
Locomotion assays and quantitative movement phenotyping.
Muscle degeneration and ageing of contractile tissue.

Deliverables What you receive with every project

Genetically verified unc-54 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, muscle- or cilia-marker, and co-CRISPR selection backgrounds.
Optional: phenotype confirmation, sarcomere and axonal imaging, and locomotion analysis support.

Frequently Asked Questions

What type of unc-54 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-54?

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-54 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.

Why does a null sometimes look weaker than expected?

Because MHC A (MYO-3) is still there. MHC A and MHC B occupy different zones of the same thick filament, so removing one does not abolish the filament outright — it produces an abnormal one. That residual structure can support a surprising amount of organisation. If your question is about filament assembly rather than motor output, build the null in combination with myo-3, and score filament length and A/B zone boundaries directly rather than relying on movement alone.

Start Your unc-54 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. MacLeod A. R., Karn J., Brenner S. Molecular analysis of the unc-54 myosin heavy-chain gene of Caenorhabditis elegans. Nature, 1981, 291(5814): 386–390.

For research use only.