PhiC31

Insertion of your transgenic constructs into a collection of defined attP target sites.

our benefits.

Experience the transformative power of PhiC31 services, seamlessly blending precision genetics, streamlined efficiency, and heightened data reliability to elevate your research to new heights.

Genetic Precision

Ensuring precise attP integration for reliable outcomes.

Data Reliability

Enhanced data reliability by minimizing the risk of unintended genetic changes.

Customized Screening

Tailor experiments with vibrant marker options.

PhiC31 services.

The PhiC31 integrase system allows a precise integration of an attB site containing transgenic construct into a defined attP target site in the Drosophila genome. This integration is mediated by the PhiC31 integrase, which is a recombinase isolated from a bacteriophage. You can select from a variety of different attP target sites.

Injection Only

Injection of your construct into the desired phiC31-attP strain and shipment of the surviving larvae. The customer is responsible for identifying the transformants. Please be aware that the PhiC31 integrase is still present in the shipped larvae.

Balanced Stocks

Currently available for 2nd and 3rd chromosome. After injection the transformant flies will be crossed with an appropriate balancer strain (2nd: yw; Gla/SM6a. 3rd: yw;; D gl3/TM3 Sb Ser). The balanced stocks will be shipped and we will maintain balanced stocks for at least 4 weeks.

F1 Transformants

Injection of your construct into the desired phiC31-attP strain, outcross of the injected males with y-w- females, and screening for transformants. Individual transformant males (F1) will be crossed with the y-w- strain and the resulting offspring will be shipped. We will maintain transformant strains for at least 4 weeks.

We can screen for the following: white, yellow, vermilion, GFP, RFP or DsRed.

the pricing.

Prices listed are for constructs <20 kb.
For constructs >20 kb we will provide a quote for you.

Injection Only

–

200 CHF

Injection of your construct into the desired phiC31-attP strain and shipment of the surviving larvae.

Processing time:

~2 weeks

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F1 Transformants

(y+ & w+)

250 CHF

Outcross of the injected animals and screening for transformants.

Processing time:

~6-7 weeks

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F1 Transformants

(vermilion & fluorescent markers)

290 CHF

Outcross of the injected animals and screening for vermilion and fluorescent markers.

Processing time:

~6-7 weeks

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Balanced Stocks

(y+ & w+)

290 CHF

Transformant flies will be crossed with an appropriate balancer strain.

Processing time:

~9-10 weeks

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Balanced Stocks

(vermilion & fluorescent markers)

330 CHF

Transformant flies will be crossed with an appropriate balancer strain.

Processing time:

~9-10 weeks

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FAQs

How much DNA should I send for the PhiC31 Injection service?

For site-specific transgenesis please provide 30 μl or more of your DNA construct at a concentration of 300 ng/μl or higher. We will dilute the DNA to an appropriate injection concentration.

If you want us to take care of the DNA you can just send us a small aliquot of miniprep DNA and we will prepare injection quality DNA in-house for CHF 30.

What happens if you don’t get any transgenics with the phiC31 service?

We offer one free re-injection if you can confirm that the plasmid is ok and should not cause lethality in a transgenic situation.

Our experience is that close to 100% of our phiC31 injections work in the first attempt. In the rare cases where we did not get transgenics in the first round the reason was mostly a problem with the injected plasmid: the plasmid is not meant for phiC31 injection (it is missing an attB recombination site); the marker is damaged or of a different nature than originally specified; there is no plasmid present (e.g. due to degradation); the transgene is e.g. an oncogene and likely causes lethality when inserted, due to leaky expression at the insertion site (leaky expression at attP target sites is a common phenomenon).

In short, a re-injection of plasmids with such issues doesn’t help to get transgenics. Therefore, we recommend to first re-check and evaluate the plasmids before doing a re-injection. We can assist you with this.

Which landing site do you recommend for site-specific integration?

With respect to integration success, fitness and survival rate we recommend: 86Fb, VK33, VK37 and VK22.