The fruit fly, Drosophila melanogaster has been an influential invertebrate model organism in understanding gene function for over a century. In addition to being suitable for classical genetics, its adaptability to evolving gene modification tools ranging from transgenesis to novel genome modification tools including CRISPR has made the fly a sought after model organism in modern biology. Since 2009 the Fly Facility, has provided services to facilitate modern molecular genetics in Drosophila.

 

 

Drosophila Transgenesis:

P-element based transgenesis: Injection of DNA into the embryos of w1118 injection stocks along with helper plasmid, crossing of G0 flies, screening the transformants and balancing the insertions.

PhiC31integrase mediated transgenesis:

Site-Specific Excel - Injection of DNA into the embryos of PhiC31 integrase & selected attP docking site (click here and choose attP sites), crossing of G0 flies, screening the transformants and balancing the insertions.

Site-Specific BAC- This service includes injecting the BACs or fosmids (>30kb) into 500 embryos of strain (or a cross of) PhiC31-integrase and a docking site of the user’s choice and scoring the transformants, balancing and shipping.

Site-Specific CRISPR- This service includes injecting the plasmids with CRISPR/Cas9 guide RNA into 300 embryos of strain (or a cross of) PhiC31-integrase and a docking site of the user’s choice and scoring the transformants, balancing and shipping.

 

CRISPR-Cas9 based genomic editing:

The fly facility uses CRISPR-Cas9 system to generate deletions.  The use of the CRISPR-Cas9 nuclease system is a very young but rapidly developing area of modern biology. The type II CRISPR system from S. pyogenes has been adapted for inducing sequence-specific DSBs and targeted genome editing. In this system, Cas9 endonuclease and a target sequence specific guide RNA (gRNA) are introduced into a heterologous system to generate DSBs in their genome in a sequence specific manner, making it a useful genetic engineering tool to induce deletions, insertions, and precisely defined modifications in genomes of diverse organisms.

This technique has been used in Drosophila to mutate >100 genes. Our facility is providing the service of sgRNA injections in various fly backgrounds in addition to full genomic deletion service. Please write to us for further details.