Optogenetics requires cell specific gene expression to manipulate neural activity in a specific group of neurons. In Drosophila, transgenic methods are widely used for this purpose.
The transgenic system mediated by phiC31 integrase is based on site-specific phage phiC31 integrase, which mediates sequence orientation, irreversible and efficient integration between bacterial attachment site (attB) and phage attachment site (attP).
Creative BioMart is now able to provide custom Drosophila strain construction service through PhiC31 Integrase-mediated Transgene Method. The service includes the injection, generation and identification of transformants. We will inject your purified attB construct into your preferred attP docking site embryo containing phiC31, and return the stable line or equilibrium line without phiC31 source according to the service you choose.
Injecting a plasmid containing attB site and white label into a docking site strain containing attP with phiC31 activity can make the generated stable W + transformant contain the gene of interest between attL and attP sites (irreversible).
According to our equipment platform, we will generate transgenic fruit flies for optogenetic research according to the following process.
Scheme 1
This is an injection only service, and the cost is based on the injection of 200 attP G0 embryos. G0 embryos will be collected from the hybridization between the docking site strain you select and the phiC31 source strain. Injected G0 larvae will be delivered to you, and you are responsible for identifying your own transformants and removing the phiC31 source (by phenotype).
Scheme 2
The service includes injection, generation and identification of transformants. We will inject your attB DNA sample into 200 embryos, return the selected surviving G0 adults, expand the G1 transformant by hybridizing again, and only deliver the G2 transformant to you.
When using this service, we will try to obtain at least one independent transformant for a reasonably sized construct (less than 13 KB), because in most cases, you only need one cell line to experiment. If more lines are found during the screening process, we will send you up to 5 strains.
Scheme 3
The service includes injection, generation, identification of transformants and balancing. In addition to option 2 services, we will also balance the transformants. We can send you up to 5 balanced transformants. For the special cases described in scheme 2 (i.e. toxic or fault conversion), we will take the same measures and charge in the same way.
Creative BioMart is always devote us to provide high-quality and satisfactory service to our customers, if you are interested in our services or have some question, please feel free to contact us or make online inquiry.
Reference
