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The nematode Caenorhabditis elegans is widely used in genetic and biomedical research. DNA fragment insertions can be introduced into the C. elegans genome. Our C. elegans knock-in services include point mutations, floxed alleles, degron tagging, fluorescent tagging and immunotagging.
In the past 5 years, InVivo Biosystems has made over 600 fluorescent transgenic lines including 15 different fluorophores, 8 of which are variants of GFP.
Fluorescent Tagging. Intestinal nuclei from late L4 worms in an lmn-1p::GFP::LMN-1 transgenic strain. The image is lmn-1 tagged endogenously with GFP. Image courtesy of Dr. André Catic, Baylor College of Medicine.
Visualize your protein via addition of a fluorescent protein tag at the native locus.
Use your protein for biochemical studies by adding an immunotag at the native locus.
The best gene editing method for creating small, precise edits to introduce a small number of nucleotide changes at a target site.
Replace a C. elegans gene with an orthologous gene from human or any other organism.
Note: Additional charge applies for cargo size above 200bp for all packages.
Our Point Mutation service uses CRISPR/Cas9, which is the best gene editing method for creating small, precise edits to introduce a small number of nucleotide changes at a target site.
With this service you can:
Our CRISPR/Cas9 Editing Is Typically Very Efficient
Among the F1 candidates with the Co-CRISPR edit, our average percentage of animals with the target edit was 65.7%.
Edit percentages as high as 95% were observed.
We Can Generate Point Mutations Quickly
For these projects, our average time from injection to sequence confirmation of two independent homozygous lines was 15.3 days.
Our quickest project only took 10 days!
96 Point Mutations In 1 Gene
All 96 mutations were created in the STXBP1 gene (which is associated with epilepsy in humans) via CRISPR.
The worm homolog of STXBP1, unc-18, causes uncoordination and near-complete lack of pharyngeal pumping when knocked out.
The functionality is restored by replacing the worm gene with the coding sequence for human STXBP1.
Fluorescent tagging of genes is widely used in many model organisms to study which tissues a gene of interest is expressed in, where in the cell a gene is expressed, or to determine whether multiple genes of interest are expressed in the same location. C. elegans is particularly well suited to fluorescence studies because they are transparent. Fluorescence can be easily observed in both live and fixed images.
Knock-in a fluorescent protein at the endogenous locus using CRISPR gene editing technology.
Adding a fluorescent protein tag at the endogenous locus enables you to:
Choose From a Variety of Fluorophores
You can choose from a variety of fluorophores including YFP, BFP, eGFP, mCherry, mOrange, mScarlet, and many more. All of our fluorescent fluorophores are optimized for expression in C. elegans via codon optimization and insertion of introns.
Use our fluorescent fluorophore tags in combination with the Fluorescent Protein RNAi to reduce or eliminate fluorophore expression.
Concerned that the addition of a fluorescent fluorophores will inhibit protein function? We can use a 2A or SL2 sequence to separate the 2 components!
Things to consider when ordering your strain:
We are not limited to a fixed list of fluorophores. If you have a fluor that you would like to use, you can provide us the sequence and we will incorporate it into the design of your transgenic project. If you need your fluorescent transgenics lines to be imaged, we can readily help you with that too! If you are unsure, our genetic engineering experts will be glad to advise you. Contact us.
We are not limited to a fixed list of fluorophores. If you have a fluor that you would like to use, you can provide us the sequence and we will incorporate it into the design of your transgenic project. If you need your fluorescent transgenics lines to be imaged, we can readily help you with that too! If you are unsure, our genetic engineering experts will be glad to advise you. Contact us.
The addition of an immunotag at the endogenous locus enables you to quantify your protein without altering the level of gene expression.
Immunotags are added using CRISPR/Cas9 gene editing technology.
You can choose from a variety of tags including FLAG, HA, HIS, TAP, or S-peptide.
All tags have been optimized for expression in C. elegans.
You can knock-in a degron tag on an endogenous protein or fluorescent protein with our degron tagging service.
Adding a degron tag enables you to:
You can choose from a variety of degron inducible and developmental systems including Auxin Inducible Degradation, Photosensitive or blue-light inducible degradation, degron-tagged reporters of membrane topology, and OMA-1 tags that work to degrade protein from the 1-cell stage.
Use our degron tags in combination with the fluorescent tagging to visualize protein expression and degradation.
Things to consider when ordering your strain:
For auxin-inducible degradation, TIR1 is necessary to generate a functional recognition complex and achieve successful target degradation. If you do not have a TIR1-expressing strain ready, we can suggest the most suitable commercially available strain, or we can generate a custom TIR1 strain for you.
If you are unsure, our genetic engineering experts will be glad to advise you. Contact us.
Whole gene humanization allows you to replace a C. elegans gene with an orthologous gene from human or any other organism. If the human gene rescues the function of the gene deleted-KO, you know there is conserved biology. This is often the first step in a project to create a model to study clinical variants.
We take care to design our humanization projects to preserve endogenous transcription signals so the expression patterns and levels will be maintained. In addition, we have a proprietary sequence optimization protocol that we use to create a transcript that will be expressed well in C. elegans.
Knock-in your human (or other animal) gene of interest at the locus of an orthologous gene using CRISPR gene editing technology.
Creating a whole gene humanized line enables you to:
Note: Additional charge applies for proteins over 2000 amino acids in size for all packages.
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An expert in CRISPR genome editing, InVivo Biosystems creates custom genome-edited C. elegans and zebrafish models to enable aging, developmental, and disease studies. Our unique in vivo platforms and technologies bridge the gap between cells and mice.
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