In the process of analyzing the project recently, Xiao Bian encountered such a problem. What about the experimental verification of a gene that drives a certain function obtained through various means (selective clearance analysis, comparative genomics analysis, transcriptome analysis)? Generally divided into: After gene up regulation (gene transfection method) and gene down regulation (gene interference method), the protein expression level, changes in cell function, and changes in related functions in animal models in vivo were observed.
I believe that some of you have such doubts. I have collected the following information in a targeted way and hope it can help you.
1、 RNA interference technology
RNA Interference（ RNA interference, RNAi ）It refers to the highly conservative, double chain RNA （ double-stranded RNA ， dsRNA ）Induced, homologous mRNA The phenomenon of efficient and specific degradation. Due to the use of RNAi Technology can specifically eliminate or turn off the expression of specific genes, so this technology has been widely used to explore gene function and the treatment of infectious diseases and malignant tumors.
In brief, if a gene drives to control the function of red flower color (for example), we use RNA interference technology to degrade the mRNA transcribed by this gene, and then look at the change of flower color. If it is not red, it indicates that this gene does have this function.
2、 Gene Knockout Techniques
Gene knockout is a new molecular biological technology developed since the late 1980s. It is a technology to inactivate or delete specific genes in the body through a certain way. In general, gene knockout mainly uses the principle of DNA homologous recombination to replace the target gene segment with the designed homologous segment, so as to achieve the purpose of gene knockout. With the development of gene knockout technology, in addition to homologous recombination, new principles and technologies have been gradually applied. The more successful ones are gene insertion mutation and iRNA, which can also achieve the goal of gene knockout.
3、 yeast two hybrid
The yeast two hybrid system is a system in which the genes of the two proteins to be studied are cloned into the DNA binding domain genes of the transcription activating factors (such as GAL4) of the yeast expression plasmid and the activation domain genes of the transcription activating factors (such as GAL4), and then constructed into fusion expression vectors. The interaction between the two proteins is analyzed from the expression products
Because direct protein interaction is an important way for proteins to play a regulatory role, if you want to study a gene driving a certain function, you can integrate this gene and the target gene into yeast for interaction research, so as to analyze its function.
4、 Other Technologies
The above information is collected from the network. I believe there should be more and more detailed technologies and methods. I hope you will give me some advice.
