Zebrafish Gene-Editing Services
Design Construct Deliver Custom Gene-Edited Zebrafish
We are a zebrafish gene-editing CRO that offer multiple gene services including but not limited to mutagenesis, tagging, and injection. We use CRISPR as our genome editing method. Our service can significantly reduce your experiment design time, and can be tailored to different experimental requirements. Over 40 years of experience makes us a good outsourcing zebrafish CRO in this field which specialized to aid more researchers in their discovery.
Using CRISPR/Cas9 technology, we can modify zebrafish genome through Knockout, Point Mutation, Precise Deletion, Floxed Allele, etc. Specific experiment requirements can be met by working with our experienced expert team members.
The Point Mutation service using zebrafish introduces a small number of nucleotide changes at a target site using CRISPR/Cas9. With this service you can: study a disease-causing mutation; humanize a critical amino acid; explore the binding site of an enzyme; introduce phosphomimetics; mutate isoform start sites or make any specific mutation of interest.
Gene knockouts are a commonly used tool for biologists to understand gene function. Examination of phenotypes when the gene is deleted can reveal insights into what role that gene plays in the organism. Using CRISPR/Cas9, we can delete integral domains or the entire the coding sequence of a gene in zebrafish, depending on gene size.
The Precise Deletion service removes a small defined region of DNA via CRISPR/Cas9 gene editing in zebrafish. This can be useful for deleting protein domains, DNA regulatory regions, or any sequence of your choice.
CRISPR/Cas9 is used to insert loxP sites flanking the region to be deleted in zebrafish. This line can then be crossed with a Cre-line expressing the Cre recombinase under a specific promoter or injected with a plasmid containing Cre recombinase. The Cre recombinase promotes recombination of the two loxP sites and the region between the sites is removed from the genome.
We can help you visualize protein expression and help you conduct biochemical studies of your gene of interest using CRISPR/Cas9. Custom tags and combinations(i.e. GFP::3xFLAG, etc) are also available.
The addition of a fluorescent protein tag at the endogenous locus enables you to visualize your protein in vivo without altering the level of gene expression. In Zebrafish, fluorescent proteins are added using CRISPR/Cas9 gene editing technology. You can choose from a variety of protein tags including YFP, BFP, GFP, and mCherry.
The addition of an epitope tag at the endogenous locus enables you to quantify your protein without altering the level of gene expression. Epitope tags in zebrafish are added using CRISPR/Cas9 gene editing technology. You can choose from a variety of tags including FLAG, HA, HIS, TAP, or S-peptide.
Obtain gene knock-ins (KIs) in zebrafish with expression at
random insertion loci via Tol2.
Insert large content in the zebrafish genome using the Tol2 transposon-based system. Using Tol2 transgenesis you can generate: Cre Recombinase-expressing lines; Transcription reporters; Models to analyze tissue-specific gene expression; Models to control the timing of gene expression; Humanized zebrafish. Insertion content can range between 2 – 10 kb.
Complete CRISPR Injection Mix
Reduce time from experiment conception to execution with the affordable Complete CRISPR Injection Mix package.
All the Power of CRISPR and More
Our Complete CRISPR Injection Mix includes Cas9 protein, an RNA-based sgRNA(s), and a donor homology construct and can be shipped as a lyophilized mix in as little as 2 weeks.Each Complete CRISPR Injection Mix project includes: consultation; sequence files for the donor homology construct and the final, edited locus; development of all reagents including the sgRNA and donor homology construct; 2 – 10 uL lyophilized injection mixes with instructions on reconstitution
We offer a variety of service packages to fit the needs and budget of your lab. Choices range from the fast and affordable Complete CRISPR Injection Mix to the comprehensive Full Build service package, allowing you to customize the components you need to accomplish your gene editing goals.
|Service||Cost||Time to ship||Shipment||Benefit|
|Injection Mix||$995 and Up||Min. 3 Weeks||Reagents||Make genome edits in yours lab-specific strains.|
|Verified Cluth|| |
$9,310 and Up
Min. 4 Months
|Injected Clutch of emboryos||Expert design and pre-screening on a budget|
26,995 and Up
Min. 9 Months
|Germline-verified fish||Receive animals ready to cross with your strains|
The NemaMetrix Workflow
NemaMetrix provides a streamlined zebrafish gene editing service. Getting customized zebrafish strain made easy.
Selection of best guide RNAs surrounding the locus to be edited.
In vivo cutting assayed by PCR/sequencing to determine the most efficient sgRNA.
Donor Homology Design
Donor homology plasmid or DNA oligonucleotide is designed based on the best sgRNA.
Injection of mix into zebrafish embryos at the
1 cell stage.
Verification of edit by PCR and/or sequencing of 10% of injected embryos.
Verification of germline inheritance, 2 week old fish
Features of Our Services
Advantageous Model Organism
Choice of Simply or Complex Build
Expert Service Team
With Over 40 Years of Experience
With Ensured Quality
Why Outsource NemaMetrix Zebrafish Gene-Editing Service
See how we can benefit your research
Reduction in Experimental Design Time
NemaMetrix help you save design time and facillitate your progress.
Custom Modification for Specific Needs
Work with us to design a workflow tailored for your requirements.
Long Years of Passion and Innovation
40 years of experience makes us the best in this field.
Our specialist are highly trained to gurantee the products and solutions are well certified
See what scientists are saying about NemaMetrix
The customer service was outstanding!
“We are still outcrossing the strain to get it ready for our screen. However, I can’t thank Knudra enough for all of their trouble-shooting and hard work to get this challenging CRISPR-based promoter swap to work!”
Organization: University at Buffalo
We’ll be looking elsewhere for our strains in the future
“The product took forever to get. It seemed like because we were a small order, we were essentially ignored and back burnered. We had to call several times to ask why it was taking…
months to do a single step in the process. By the time we finally received what we had ordered, others had outpublished us because they developed it in house. When we did receive the strain we ordered, it was clearly messed up and completely unusable except as a scientific curiosity. $3,000 of precious research money down the drain!!!”
Organization: University of Tulsa
They are a fantastic resource for the worm field.
“Setting up injections is pretty straightforward and the time of return to get back the injected animals is very quick. There was a substantial increase in price per injection which…
came as a surprise which really makes it more difficult to purchase the service for multiple injections. Now that CRISPR is becoming popular the service recommends performing 3 times as much injections to obtain the completed edit, but did not specify in the communication if this also results in 3x the cost as a normal round of injections with user generated plasmids.”
Organization: University of South Florida
CRISPR made easy!
“Super helpful in helping design and generate CRISPR knockouts. Communicative and transparent at all stages of the process.”
Organization: Worcester Polytechnic Institute
Great reproducible results, love the service.
“Amazing service and terrific support. The strain I ordered behave as expected, is ready to use and produces reproducible results.”
Great results! So glad I used this time saving service.
“I requested a new C. elegans line generated by knocking out a second gene in animals with a deletion…
in a second, highly homologous, closely linked gene. The design and screening of this was very difficult for me in my own laboratory, and Knudra had trouble generating the line. This alone made it worth the money that it took for them to do it because at the end of the day they delivered three lines that seem to have consistent phenotypes with what i expected. I loved that they did the designing of the guide RNAs and everything else to generate this knockout, that they communicated with me throughout the process, and that, when one of the three lines was not behaving properly, they went back to find the correct phenotype in animal stocks that they kept.”
Organization: Georgia state university
Really great service – all of the mutations that we needed were generated
“We ordered a series of gene edited worm strains for genes that we are interested in relating to signal transduction. The ordering and design process was straightforward, and we were kept up to date with the generation of the mutants and any…
to date with the generation of the mutants and any potential obstacles. All of the mutants we ordered were generated and delivered, very impressed with the speed and quality of service.”
Organization: University of Reading
Reluctant to order another mutant from Knudra in the future because of loss of time
“Service was rapid but the delivered homozygous mutant was not a mutant. Upon complaint, Knudra stated that homozygous mutants were not stable in their hands suspected lethality of homozygote and recommended suspected lethality of homozygote…
suspected lethality of homozygote and recommended crossing heterozygotes over a balancer strain. New injections were made and heterozygotes delivered to customer, which we crossed over balancer strains. However, it turns out the heterozygotes can be bred homozygously without need for balancer strain. In conclusion, we now have what we wanted but lost several months of time. I wish the strain had been checked more carefully by Knudra before sending the first time.”
Organization: University of Lausanne
Very good result, will certainly use the service again when needed
“Knudra provided a fast, very professionnal handling of our project. This is a difficult project implying many steps and Knudra provided efficient feedback at all stages. The final delivery was just fine. I appreciated the customer-directed approach with minimal paperwork.”
Organization:The University of Namur
See how NemaMetrix is helping scientists with zebrafish gene editing
Mutagenesis in EYS Gene
Mutations in EYS are known to cause photoreceptor degeneration in patients with RP25, but functions of the EYS protein in the vertebrate retina are poorly understood.
ABC research lab aims to study the function of Eye shut homolog for survival of photoreceptors in zebrafish. They proposed several method of mutagenesis in EYS and found out that CRISPR/Cas9 is the best technique. However, due to lack of experience in practicing CRISPR/Cas9, ABC researchers had difficult time introducing mutations of EYS gene on the cone photoreceptors. After several unsuccessful trials, ABC researchers have still not found out a functional protocol to knockout EYS gene.
While searching for the protocols for zebrafish CRO to outsource, ABC researchers discovered the zebrafish gene-editing service offered by NemaMetrix. NemaMetrix zebrafish service team planned out a detailed workflow for zebrafish EYS gene knockout, and carried out the entire project in a turn-key fashion. After 6 months, the genetically modified zebrafish was delivered to the lab, after which observations were made every week on the development of retinal photoreceptors. A deficiency of EYS protein was observed in all 200 individuals, suggesting a 100% knockout success rate. Progressive degeneration of cones and rods were discovered to correlate with decreased level of EYS protein. ABC derived a solid conclusion from this research and published paper after confirming their prediction with the help of NemaMetrix’s zebrafish gene editing service.
Measure Expression Level by GFP
B pharmaceutical company designed a drug de novo aiming to treat Reticular Dysgenesis and advanced to in vivo testing of the de novo drug in model organisms. After intense debate on which model organisms should be chosen, zebrafish was determined to be used considering its relative advantage to model human diseases especially pediatric genetic diseases.
However, B company is not experienced in genetically modifying zebrafish genome, so the research team decides to seek for assistance from a zebrafish CRO that specialize in gene editing. They browsed several zebrafish CROs and finally decided on outsource NemaMetrix due to its experience and credential. After discussion about this case, both companies decided on using GFP tagging to indicate the expression level of AK2 gene before and after usage of de novo drug, and different dosages were applied to discover the optimum daily usage. Level of expression were determined by the measuring the amount of fluorescence. The drug was discovered to be effective in raising the expression of AK2 proteins, indicted by the increasing level of GFP detected after drug injection. B company was able to proceed to the next stage of testing drug side effect.
Introduction of Tissue-Specific KRAS Expression
A zebrafish research lab in University of C hoped to figure out the differential expression of a oncogenic gene KRAS in different types of tissue. A main focus of this experiment is to see how the nervous system can be affected by KRAS mutation. Researchers have already decided that Cre-lox system would be a best fit in this scenario. Due to limited knowledge of zebrafish gene editing, the researchers sought assistance from other zebrafish research labs, during which they obtained information about several zebrafish CROs, and finally agreed to outsource NemaMetrix. They were interested by the TOL2 transposon-based system as well as the efficiency service delivery. Out of three options, verified was chosen due to short preparation period. The modified zebrafish were delivered three months after the order, and subsequent experiment was successful. Using Cre-Recombinase inducing drugs, tissue specific loss of function mutation of KRAS was achieved by the researchers. Observations were made and they drafted a paper describing their findings.
CRISPR Based Incoporation of THOR gene
D lab was working on discovering cancer promoting genes. They managed to target a human gene named THOR which facilitate the growth of melanoma in human. In order to validate their hypothesis, researchers in this lab picked zebrafish as an experiment model. Zebrafish is known to be an advantageous model to study human disease. The team concentrated on discovering human oncologic genes, and was not able to spend too much time on the validation process, especially on reagent design and validation. However, as a crucial part of the experiment, D lab cannot derive a conclusion without testing their result. D lab did a research online and found out that NemaMetrix had a suitable option for them to outsource which is the complete CRISPR injection mix. With this option, they only need to provide NemaMetrix with the THOR gene construct, and the injection mix could be shipped back to them in as short as 2 weeks, including Cas9 protein, an RNA-based sgRNA(s), and a donor homology construct as a lyophilized mix. Lab D started the process right way and received the injection mix in two weeks. They then injected the mix into 100 different zebrafish, which are chemically induced to develop melanoma. They found out that THOR incorporation into the zebrafish genome induced faster tumor growth in the model, and thus proved their hypothesis to be correct.
CRISPR Based Incoporation of THOR gene
Lab E in XY medical school specialize in the research of drugs to target Cystic Fibrosis’s symptoms. A drug testing on model organism was needed before clinical trials on human could be done. Researchers in lab E favors zebrafish because it has a genome that is easy to work with. However, they have nearly zero experience working with zebrafish. They planned to introduce a functional copy of CFTR gene into one group of zebrafish and one point-mutated copy of CFTR into another group of zebrafish. After weeks of struggle, they could not produce viable zebrafish with precise point mutation on a donor copy. As they were about to start another trial, they were informed by their partner lab that NemaMetrix is a gene-editing company which specializes in zebrafish point mutation. They immediately outsourced NemaMetrix and explained their situation, after which NemaMetrix offered a custom service. Two groups of zebrafish as desired were shipped to Lab E after 6 months. And Lab E was able to complete the do a comparative study to test the effect of point mutation on effective CFTR gene.