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CN-121987829-A - Application of RNA-SNORA A in preparation of medicine for treating pancreatic cancer

CN121987829ACN 121987829 ACN121987829 ACN 121987829ACN-121987829-A

Abstract

The application provides an application of RNA-SNORA A in preparing a medicine for treating pancreatic cancer, wherein SNORA A is used as a medicine target for treating pancreatic cancer, and the gene sequence of SNORA A is SEQ ID NO.1.

Inventors

  • CUI CHUNHONG
  • CHEN YUWEI
  • CHEN WEILING
  • WANG HUI

Assignees

  • 上海健康医学院

Dates

Publication Date
20260508
Application Date
20260403

Claims (7)

  1. 1. An application of RNA-SNORA A in preparing a medicament for treating pancreatic cancer, which is characterized in that the gene sequence of SNORA A is SEQ ID NO.1.
  2. 2. The use of RNA-SNORA a according to claim 1 for the preparation of a medicament for the treatment of pancreatic cancer, wherein tumor cell invasion is inhibited by reducing pancreatic cancer cell neogenesis protein synthesis by knocking out SNORA a in the adenocarcinoma cells.
  3. 3. The use of RNA-SNORA a according to claim 1 or 2 in the preparation of a medicament for the treatment of pancreatic cancer, characterized in that a plasmid vector constructed from the gRNA shown in SEQ ID No.2 and Cas9 protein is transfected into pancreatic cancer cells to be knocked out based on CRISPR/Cas9 technology, and the pancreatic cancer cell line knocked out SNORA a gene is obtained by screening.
  4. 4. The use of RNA-SNORA A as defined in claim 3 for the preparation of a medicament for the treatment of pancreatic cancer, wherein the guide sequences of SNORA A are SEQ ID NO.2 and SEQ ID NO.3.
  5. 5. The use of RNA-SNORA a according to claim 4 for the preparation of a medicament for the treatment of pancreatic cancer, wherein the plasmid vector is one of pX330, pSpCas9 (BB) -2A-GFP, pLenti-CRISPR v2, pCRISPR/SA, pX459 or pL-CRISPR. Efs. GFP.
  6. 6. The use of RNA-SNORA a according to claim 4 for the preparation of a medicament for the treatment of pancreatic cancer, wherein the construction step of a plasmid vector comprises: Cutting the amplified Cas9 gene and grnas shown in SEQ ID No.2 and SEQ ID No.3 using a restriction enzyme, and the plasmid vector; And (3) connecting the annealed gRNA sequence with the plasmid vector under the action of DNA ligase.
  7. 7. The use of RNA-SNORA a according to claim 1 in the manufacture of a medicament for the treatment of pancreatic cancer, wherein the screening method uses one of antibiotic screening, flow cytometry, single cell sequencing.

Description

Application of RNA-SNORA A in preparation of medicine for treating pancreatic cancer Technical Field The application relates to the technical field of bioengineering, and further relates to application of RNA-SNORA A in preparation of a medicine for treating pancreatic cancer. Background Pancreatic cancer is a highly malignant tumor characterized by early diagnosis difficulties, rapid progression and limited treatment options. The occurrence and progression of pancreatic cancer involves mutations in multiple genes and abnormal activation of signaling pathways. Currently, the treatment of pancreatic cancer mainly includes surgical excision, chemotherapy, radiotherapy, targeted therapy, etc., but the overall prognosis is still poor. In recent years, researchers have made significant progress in the genetics and molecular biology of pancreatic cancer. For example, mutations in the K-ras gene are very common in pancreatic cancer and are considered an early event in pancreatic carcinogenesis. In addition, mutations in genes such as CDKN2A (encoding p16 protein), TP53, SMAD4, and BRCA2 are also closely related to the occurrence and progression of pancreatic cancer. Gene knockout technology is an important tool for studying gene function. Traditional gene knockout methods include homologous recombination mediated gene knockout and RNA interference (RNAi) techniques. However, these methods have some limitations such as inefficiency, long cycle time, off-target effects, etc. The advent of current CRISPR/Cas9 technology has greatly driven the development of the field of gene knockout. CRISPR/Cas9 technology utilizes a stretch of guide RNA (gRNA) to guide Cas9 nuclease to specific gene locations for cleavage, thereby achieving efficient gene knockout. Compared with the traditional gene knockout method, the CRISPR/Cas9 technology has higher efficiency, specificity and flexibility. Patent CN110343700a discloses a pancreatic cancer cell line with XPO1 gene knockout and a construction method thereof, the XPO1 gene of the pancreatic cancer cell is knocked out by using a gene knockout technology, and the nuclear transport mechanism of XPO1 targeted by abnormal indication in various malignant tumors can become a potential therapeutic target of the malignant tumors. Aiming at the potential target point of XPO1, the patent establishes a pancreatic cancer cell line capable of realizing stable inhibition of XPO1 genes at a genome level, and provides an important tool for related research. The research on potential gene therapy targets in the field of pancreatic cancer therapy still has been insufficient, so the application aims at finding out another research path of potential gene targets, provides a new technical strategy for research on pathogenesis of pancreatic cancer and development of therapeutic means, and can powerfully promote the progress of pancreatic cancer related research. Disclosure of Invention Aiming at the problem that the research on potential gene therapy targets by the current pancreatic cancer treatment means is insufficient, the application aims to provide an application of RNA-SNORA A in preparing a medicament for treating pancreatic cancer, wherein RNA-SNORA A is used as a target for treating pancreatic cancer, and the gene sequence of SNORA A is SEQ ID NO.1. According to one embodiment of the application, tumor cell invasion is inhibited by knocking-out SNORA a from adenocarcinoma cells to reduce pancreatic cancer cell neogenesis protein synthesis. According to one embodiment of the application, based on CRISPR/Cas9 technology, a plasmid vector constructed by gRNA shown in SEQ ID NO.2 and Cas9 protein is transfected into pancreatic cancer cells to be knocked out, and the pancreatic cancer cell line knocked out SNORA A gene is obtained through screening. According to one embodiment of the application, the guide sequences SNORA A are EQ ID No.2 and SEQ ID No.3. According to an embodiment of the application, the plasmid vector is one of pX330, pSpCas9 (BB) -2A-GFP, pLenti-CRISPR v2, pCRISPR/SA, pX459 or pL-CRISPR. EFS. GFP; According to an embodiment of the present application, the construction step of the plasmid vector includes cleaving the amplified Cas9 gene and gRNAs shown in SEQ ID No.2 and SEQ ID No.3 using restriction enzymes, and the plasmid vector; And connecting the annealed Cas9 gene and the gRNA sequence with the plasmid vector under the action of DNA ligase. According to an embodiment of the present application, the screening method uses one of antibiotic screening, flow cytometry, single cell sequencing. Compared with the prior art, the application has the following beneficial effects: According to the application, SNORA A gene is established as a medicine treatment target point of pancreatic cancer for the first time, CRISPR/Cas9 gene knockout technology is utilized to accurately knockout the gene from pancreatic cancer cells, and a plurality of experimental results show that after SNORA A