Biological recombinant miR124-3p capable of effectively suppressing osteosarcoma growth

Abstract

Biological recombinant miR124-3p capable of effectively suppressing osteosarcoma growth is disclosed and has a nucleotide sequence shown as SEQ ID NO:1. A base sequence capable of expressing mature miR124-3p is inserted from an anticodon in the base sequence of human-derived tRNA adopted as a carrier skeleton, then the synthesized structure is transferred into common E. coli through a plasmid, andafter about 12 h of incubation, a target recombinant miRNA is subjected to separation, purification and desalting treatment to obtain the recombinant miRNA that can be put into use directly. Under the premise of ensuring a high yield and high purity, the whole preparation process adopts natural organisms as media, and original biological characteristics of natural miR124-3p in cells are preservedwith extremely high fidelity, and therefore, the miR124-3p is safer, stabler and more natural than miRNA analogues artificially chemically synthesized.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to a novel recombinant miR124-3p, i.e. htRNA, which can be constructed and produced on a large scale by genetic engineering methods Leu / miR124-3p. Background technique [0002] Osteosarcoma (OS) is the most common primary sarcoma in children and adolescents, and the incidence rate of males is higher than that of females, with a ratio of about 2:1. From an anatomical point of view, the most common sites of osteosarcoma are the proximal end of the distal femur, tibia, and humerus, and about 50% to 70% of all patients occur around the knee joint. At present, in clinic, the main treatment for osteosarcoma is neoadjuvant chemotherapy---resection of local tumor tissue by surgery---maintenance of chemotherapeutic drugs. It has greatly reduced the amputation rate of patients with affected limbs, and significantly increased the five-year survival rate of patients from about 20% ...

AI Technical Summary

Problems solved by technology

[0004] With the deepening of non-coding RNA research and understanding, the important role of miRNA molecules in new cancer treatment schemes is becoming more and more obvious. By referring to a large number of literatures, we can clearly find that no matter what Whether it is in the laboratory research stage or in the clinical practice, the vast majority of miRNAs used by researchers are artificially synthesized, while the miRNA reagents obtained by other methods are very small and very limited.

As we all know, miRNAs synthesized by artificial chemical methods need to undergo a large number of special artificial group modifications to ensure the stability of their structures and biological functions. However, as manufacturers add artificial chemical groups of different structures to miRNA analogs, , the side effects of these artificial groups themselves also appear, and how much these artificial modification groups can specifically bring to the biological characteristics of miRNA itself, such as the multidimensional structure, physical and chemical properties, biological activity and biological safety of miRNA, etc. At present, there is no suitable method and effective way to accurately detect and express the influence and change. Therefore, these artificially modified chemical groups will definitely have a non-negligible impact on researchers in the process of studying miRNA

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