Genome engineering as an approach to identify crucial miRNA targets in cancer

All the cells composing the body contain the genetic information necessary for the life of each individual. Genetic information is encoded by the deoxyribonucleic acid (DNA), which represents the genomes and exerts its function via making proteins. A crucial intermediate in this process is ribonucleic acid (RNA) which is specifically called messenger RNA (mRNA). Recently, it has been shown that besides mRNA, RNA that does not produce proteins exists and its production and function in the cells are equally important to the ones employed by mRNA. Because the latter class of RNA is not created by the cells to produce proteins, but instead to have a regulative role, it is called noncoding RNA (ncRNA). Similarly to mRNA, the production and function of ncRNA ensure an accurate cellular function and consequently a healthy life. NcRNA role is to precisely regulate mRNAs when they produce proteins. This means that the production of proteins by mRNA can increase or slow down depending of particular circumstances, and this effect is mediated by the interaction between mRNA and ncRNAs. NcRNAs can be divided into different classes, depending on their length. MicroRNAs belong to the short ncRNAs class and several studies performed on microRNAs, in the last decade, have demonstrated that these short RNA molecules are important regulator of cancer development. Moreover, specific drugs can now be developed to specifically target microRNAs in the body to fight cancer or other diseases. MicroRNA function by interacting with small, specific region of mRNA to slow down protein production. In the last few years, other important discoveries, in science, are remarkably impacting our understanding of the way that the cells function as well as about what goes wrong when the cells pathologically change and form cancers. One of these new techniques is called CRISPR-CAS9 and can be used, in vitro and in vivo, to precisely edit the DNA of the cells accordingly with our needs. Because DNA generate RNA the new modification created at the level of DNA, by using CRISPR-CAS9 in the cells, will be consequently transferred to the RNA. Herein, we want to use CRISPR-CAS9 to globally disrupt the mRNA portions predicted to interact with microRNAs. Because disruption of these mRNA regions will impede functional microRNA-mRNA interaction, the mRNA will consequently produce a higher amount of protein that can be deleterious and induce cancer. In principle, mRNA that produce proteins important for cancer, can also be modulated by microRNAs. By performing specific laboratory tests and by studying changes in the growth of tumour cells seeded on specific dishes and treated with CRISPR-CAS9 we can finally identify important microRNA-mRNA interactions that can be exploited for future therapeutic intervention. By using both experimental techniques and computational approaches, this work would permit us to understand the important mRNA-microRNA interactions that regulate cancer development and this new knowledge could then be used to develop new drugs to fight cancer.