Lung cancer fusion gene nucleic acid detection quality control product based on CRSIPR-Cas9 technology and preparation method thereof

Abstract

The invention provides a lung cancer fusion gene nucleic acid detection quality control product based on a CRSIPR-Cas9 technology. The lung cancer fusion gene nucleic acid detection quality control product is used for comprehensively monitoring a fusion gene detection process. A preparation method comprises the following steps of: (1) constructing a monoclonal HBE cell strain with specific fusionsites of EML4-ALK and CD74-ROS1 fusion genes by utilizing a CRISPR-Cas9 knock-in technology; (2) constructing a mouse subcutaneous transplantation tumor model by combining the monoclonal HBE cell strain successfully edited in the step (1) with a control wild cell strain; killing the model mouse after the tumor grows to 1cm<3>, collecting tumor tissues, pretreating, fixing, and embedding with paraffin; and (3) constructing a case tissue slice microarray according to a certain mutation / wild ratio by using the tumor tissues collected in the step (2) to obtain lung cancer fusion gene nucleic aciddetection quality control products with different fusion frequencies. The quality control product prepared by the invention has high similarity with non-small cell lung cancer, can truly realize the whole process of lung cancer sample tissue from sampling to detection, and realizes comprehensive monitoring of fusion gene detection.

Description

technical field [0001] The invention belongs to the field of detection of lung cancer fusion gene nucleic acid, and in particular relates to a quality control product for detection of lung cancer fusion gene nucleic acid based on tissue microarray and CRSIPR-Cas9 technology and a preparation method thereof. Background technique [0002] Lung cancer is the malignant tumor with the highest incidence rate in my country, and it is also the most important type of tumor that causes tumor-related deaths. It is predicted that by 2025, new cases of lung cancer in my country will exceed 1 million per year. Therefore, the diagnosis and treatment of lung cancer has become an imminent "tough battle" in the field of cancer research in my country. Non-small cell lung cancer (NSCLC) is the most common type of lung cancer, accounting for more than 85% of all lung cancers. Existing studies have shown that ROS1 and ALK fusion genes are new driver genes in the Chinese NSCLC population, accoun...

AI Technical Summary

Problems solved by technology

Although the previous artificial preparation of lung cancer tissue has solved most of the disadvantages of patient-derived quality control products, there are still many shortcomings: First, most of the tissue sources are tool cells such as 293T, which are not cancer cells themselves. There are a large number of differences between the transcriptome and the actual lung cancer cells; secondly, the fusion gene incidence of quality control products varies widely, especially at low abundance levels; finally, the overall process is relatively complicated, and it is difficult to transfer to actual large-scale industrial production

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