Drosophila CRISPR Transformation Services
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. We are now able to provide custom Drosophila strain construction service through CRISPR Transformation Method.
Creative BioMart is committed to providing professional services for researchers to produce transgenes and gene knockout / knockout in Drosophila models (Drosophila melanogaster). Each project will be designed, implemented and supervised by postdoctoral scientists to ensure the quality and reliability of the whole experimental process. Just let us know the information of the gene and how you want to edit it. Creative BioMart will propose the best strategy and complete the remaining steps for our customers.
Technology Principles
In the application of CRISPR / cas9 system in Drosophila genome editing, two kinds of RNA that play a key role can be synthesized into a single chimeric RNA (chirna or gRNA). By editing the 20 NT targeted sequence of gRNA, the system introduced DSB on the target sequence, followed by NGG or nag prototype spacer adjacent motif (PAM) required for cas9 cleavage.
CRISPR / cas9 works through RNA-DNA recognition, so cas9 can work simultaneously with multiple gRNA target sites, so as to promote precise deletion, insertion and sequence replacement between spaces.
Available Schemes at Creative BioMart
According to our equipment platform, we will generate transgenic fruit flies for optogenetic research according to the following process.
Scheme 1— Injection only
This is an injection only service, and the cost is based on the injection of 300 embryos. Due to the nature of cas9 lines and / or injection mixtures, we may inject more than 300 embryos in most cases. Please send us your CRISPR injection mixture (DNA, RNA, oligonucleotides, etc.) or provide us with very clear instructions on how to make the injection mixture.
Scheme 2— Injection, generation and identification
The service includes injection, generation and identification of CRISPR HDR transformants with visible markers. We will inject your CRISPR mixture into 300 embryos, return the selected surviving G0 adults to the appropriate original species, expand G1 transformants by re hybridizing to the appropriate original species, and only deliver G2 CRISPR HDR transformants to you.
Scheme 3— Injection, generation, transformant identification and balance
The service includes injection, generation, transformant identification (with visible markers) and balance. We will also balance CRISPR HDR transformants. In some cases, we can use the balancer to set G0 crossover directly.
Optional Services
The screened positive lines were isogenic and balanced. We build flight inventory for you.
- Plasmid Construction Service (<3kb)
Up to 3 PCR fragments with a total length of less than 3KB. We used high fidelity phusion DNA polymerase to amplify fragments and sequences through all parts of our clones. We returned the bacterial scribe plate and 4 µ g plasmid DNA.
Plasmid preparation to provide high-quality DNA for microinjection. Including restriction enzyme digestion or PCR validation services.
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.
