Figure1: Strategic planning flowchart (Gratz SJ, 2015) Transgenic and gene editing technology is an essential tool to study the structure, function and regulation of Drosophila gene. The use of transgene and gene editing can realize gene knockout or rescue, up-regulation or down-regulation of expression, track the period and location of gene expression, conditionally (specific time and space) express the target gene or operate the cells of specific groups, and so on.
Creative BioMart has comprehensive, mature and efficient gene editing technical services: transposase mediated random insertion transgene, integrase phiC31 mediated site-specific insertion transgene and CRISPR / cas9 gene editing, covering the whole process from scheme design to obtaining stable strains.
| Services and Technologies | Technical Features | Applications | |
|---|---|---|---|
| Transposase | PiggyBac | The insertion position is random, the phenotype is unstable, and there is a risk of damaging endogenous genes. | Differential expression of foreign genes. |
| Random insertion | P-element | Enhancer Trap. | |
| Site directed insertion of integrase | attP-PhiC31 | Foreign sequences were inserted into specific attP sites in specific Drosophila genome. | Internal and external gene expression. RNA interference. Study on the function of regulatory sequence. |
| CRISPR | CRISPR-indel | Small fragment insertion deletion, strong applicability, may produce unknown functional proteins. | Endogenous gene frameshift mutation. |
| CRISPR-KO | Large fragment knockout has certain limitations, and the knockout results are relatively stable. | Endogenous DNA sequence knockout | |
| CRISPR-KI | Insert foreign sequences into the designated target to accurately modify the genome. | Analysis of endogenous gene expression pattern. Endogenous gene point mutation, endogenous gene knockout replacement. | |
| CKOAKI(CRISPR-KO-attP-KI) | In the two-step method, the attpg replacement line is obtained first, and then the target insertion sequence is supplemented. | Targeted solutions to complex needs. | |
Overall Customization of the Whole Service Process
Figure2: The CRISPR/Cas9 system in Drosophila. (Sun, M., 2015) For the whole process packaging and customization service, Creative BioMart has taken certain measures and formulated corresponding terms to ensure the project cycle and success rate, so that customers can obtain Drosophila strains that fully meet the research purpose with less energy, shorter time and lower cost.
The customer only needs to provide the species, gene name or sequence and cloning method, and clarify the needs, we can formulate the most suitable whole process scheme, and provide the electronic version scheme (including cloning primer, vector sequence, gRNA design, etc.) for the customer's confirmation, so as to ensure the accurate realization of the customer's research purpose.
According to the needs of the project, endogenous expression vectors of various enzymes such as piggyBac, P-element > fhic31 and cas9 are provided free of charge for microinjection.
It has various types of plasmid vectors, and can provide DNA sample preparation services such as vector construction, endotoxic Plasmid Extraction and plasmid purification.
For CRISPR, the target sequence was first sequenced and confirmed, and then gRNA and cas9mrna were synthesized in vitro by self-developed method to obtain high-efficiency RNA samples.
It has an injection team with many years of experience and equipped with the industry's top microinjection instrument to ensure the continuous and stable efficiency of gene editing at a high level.
A variety of injected Drosophila melanogaster are available, including wild-type with different backgrounds, endogenous expression of cas9 Drosophila melanogaster with different chromosomes and more than 10 attP Drosophila melanogaster strains.
By transferring the screening marker gene at the same time, it is convenient to obtain the positive.
Molecular identification of screening positive Drosophila melanogaster was carried out to ensure the accuracy of genome sequence. The perfect hybridization molecular identification process obtained the positive items without screening markers.
It can be hybridized and delivered according to customer requirements, including marker removed lines, homozygous lines, multi balanced sub lines, multi chromosome stable lines, etc.
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.
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