sandramueller

Photo: Sandra Mueller

The Drosophila Injection Service provides EMBL scientists with the equipment and the expertise necessary to inject Drosophila embryos with DNA, RNA, proteins, dyes and lipids.

DNA injection

P-element mediated germ line transformation

Plasmid DNA containing a transgenic construct flanked by P-element inverted repeats is mixed with a P-element transposase-carrying helper plasmid (provided by the service) and injected into around 200 embryos of w1118 flies. Please fill in the P-element form for each DNA sample you would like to inject and prepare the DNA following the protocol for DNA preparation for P-element.

PhiC31 integrase-mediated site-specific transgenesis

This method utilises the recombination machinery of the phiC31 integrase. The plasmid, containing the gene of interest flanked by attB sites, is inserted into a predetermined attP docking site on a specific chromosome in Drosophila attP landing site lines. We provide the empty plasmid to clone your gene of interest as well as a selection of attP landing sites on the X, II and III chromosome. Please fill in the integrase form for each DNA sample you would like to inject and prepare the DNA following the protocol for DNA preparation for integrase.

CRISPR genome editing

The CRISPR/Cas9 system is a powerful genome-engineering tool that allows deletion and insertion of genes of interest at a specific locus. We provide flies carrying cas9 on the III chromosome under the control of vasa regulatory sequences (w[1118];PBac{y[+mDint2]=vas-Cas9}; credit Kate O’Connor-Giles; BDSC #51324). Around 200 embryos will be injected with a mix of the donor plasmid and the guide RNA (gRNA). Please fill in the CRISPR/Cas9 form for each DNA sample you would like to inject and prepare the DNA following the protocol for DNA preparation for CRISPR/Cas9.

In room 554, on the top of “Drosophila Injection Service” freezer in “P-Element”, “Integrase” and “CRISPR/Cas9” boxes, you will find the relevant submission forms. On the reverse side of each form you will find a protocol for DNA sample preparation, or you can download it from the links above. Please be advised that following this protocol is crucial for efficient transformation.

All tubes with DNA samples, clearly labeled with a job name, should be deposited in drawer #4 of the “Drosophila Injection Service” freezer in room 554. The submission form can either be sent by email or left in the “New Order” box on top of the freezer. Of note: job names on sample tubes and submission forms must be identical!

When the injected embryos hatch, you will receive a notification email to pick up tube(s) with hatched larvae. After you finish screening for transformants, please remember to send back the form you received with the larvae, filled in all its parts. Your feedback is very important to keep the service running optimally.

Protein, dye and lipid injections

Please contact me as soon as you start planning an experiment. I will provide you with any information needed and gladly help you to ensure efficient completion of your project.

Training to learn how to inject can also be provided.

Contact

Alessandra Reversi
Ephrussi Lab, room 542
Developmental Biology Unit
Tel: +49 6221 387-8363 or -8270
alessandra.reversi@embl.de

Please check availability of the service in the calendar (EMBL users only - please log in to your EMBL Google account first).

Please do not hesitate to contact me for any further information.