Small Interference RNA Knocks down the Expression of Gene CCP22

TGF β-Smad had wide signaling pathways involved in the regulation of various life processes, such as cell growth, differentiation, apoptosis, extracellular matrix formation and development. In these processes, Smad4 played a core role. Research shows tumorigenesis is associated with Smad4.

The domain (CCPmodule) of Complement control protein (CCP) was first discovered in 1987. There are two categories of proteins with CCP domain, regulators of complement activation (RCA) and non-regulators of complement activation (non – RCA). The CCP domain has an important function which can be combined with antigen molecules involved in cell signal recognition.

CCP22 is a new and not reported gene gained by using yeast two hybrid screening from DNA Library of human breast interacted with Smad4. The full-length of CCP22 gene is 4494bp, encoding a protein of 1497 amino acid residues. With existing data combined bioinformatics analysis, it is found that the protein encoded by this gene contains 21 CCP elements and 2.5 CCP sequence elements. So the gene was named CCP22.

 

Protein with different CCP domains, its domains are relatively conservative. But there are some differences in the number of them. Research shows that the proteins with CCP domains may also contain a transmembrane domain, receptor binding domain and other domains. Unlike other proteins with CCP domains, CCP22 only has a CCP domain. So in the CCP family of proteins, it is not clear about its ownership. The study on CCP22 contributes to the understanding of Smad4. It is possible to associate tumor development with immune complement, providing new molecular targets for drug therapy. To this end, small Interference RNA Knockdown is constructed. Western blot shows that siRNA can effectively reduce the expression of CCP22 protein.

At present, there are 3 main ways to get siRNA, chemical synthesis, in vitro transcription and siRNA recombinant expression vector construction. PSilencer2 is choosing as the siRNA expression vector. 1－U6－neo has RNApolIII promoter, and has the efficient, cheap and convenient advantages. The FlagCCP22 recombinant expression vector and CCP22siRNA recombinant vector are transfected into HEK293T cell line. Western blot shows that CCP22siRNA can effectively reduce the protein expression level of FlagCCP22. At the same time, CCP22siRNA can also reduce the endogenous CCP22 protein expression level, to lay a solid foundation for further study on the function of CCP22 protein.

 

Contact:

45-1 Ramsey Road, Shirley, NY 11967, USA

Tel: + 1-631-626-9181

Email: info@Creative-Animodel.com

Website: http://www.creative-animodel.com

 

RNA Interference for Gene Knockdown and Silence

RNA interference (RNAi) is the process by which the expression of a target gene can be effectively silenced or knocked down. RNA interference involves two types of small ribonucleic acid, namely, microRNA (miRNA) and small interfering RNA (siRNA), which can bind to specific messenger RNA (mRNA) molecules and increase or decrease their activity.

RNAi is a latest powerful gene knockdown technique that allows for tissue-specific, temporally controlled suppression of gene expression. Gene knockdown is achieved by causing the destruction of specific mRNA molecules by introducing small double-stranded interfering RNAs (dsRNA) into the cytoplasm. When siRNA is introduced into the cytoplasm of cells, it will be cut into short interfering RNAs (siRNAs) which can target specific mRNA and causing degradation of it. Therefore, the expression of a particular gene can be suppressed by introducing dsRNA whose antisense strand sequence matches the mRNA sequence. Gene knockdown by RNAi is temporary, but this can be solved by transfecting target tissues or cell lines so that they express dsRNA autonomously.

The versatility of the technique leads to a number of applications. RNAi can be used in drug or chemical target validation. RNAi can target specific spliced exons, enabling the investigation of the functional roles of alternatively spliced forms of a gene. RNAi is also valuable research tool in cell culture and in living organisms where it can be used for large-scale screens that systematically shut down each gene in the cell, thus helping to identify the components necessary for a particular cellular process or an event.

 

The discovery of RNA interference brings significant advances in both basic research and translational applications. With the elucidation of the RNA interference pathway and the discovery of its participation in crucial biological events, a branch of science has come into being to utilize the RNA interference pathway as a biotechnology for both basic and applied research. To date, small interference RNA, plasmid-, and virus-encoded short-hairpin RNA are regular reagents in laboratories to knockdown the expression of specific genes post-transcriptionally.

RNAi Gene Knockdown Services by Creative Animodel

Creative Animodel takes up long-time challenges to develop therapies for chronic and long-time illness. The company is an experience life science CRO and offer accelerated preclinical services, by using a number of bio-techniques to know more about the disease and how to combat it. RNAi Gene knockdown experiments have become very popular in the world of medical science. It has become and common and routine function to assess and study the gene functions in a quick and easy way. A very widely used approach in performing gene knockdown experiments is the RNA interference. The process is to conduct experiments on cell cultures and transgenic animals.

 

 

Creating mice with down regulated target genes involves a number of purposes. The rapid generation of the mice is done by the transduction of early embryos with adenoviral vectors and lent viral silencing. Such methodologies open up chances of large scale generation of knockdown transgenic mice, which can be used for functional genomics studies. Within just 7 seven weeks, this process enables the production of transgenic mice.

 

RNAi gene knockdown is a method which is being employed by scientists in different parts of the globe, and is yielding fruitful results. Creative Animodel, conducts all these experiments in absolute propriety. Their RNAi technology can help you in inducing and reversing ubiquitous gene knockdown in mice at different given times. There are customer services, will be happy to help you and answer all your queries in detail. You can give them a call at any time of the day and any day of the week.