SiRNA for targeted inhibition of SOS1 gene expression and application of siRNA

A gene expression and targeting technology, applied in the field of siRNA, can solve problems such as drug resistance of patients, and achieve the effect of inhibiting growth, overcoming drug resistance, and increasing drug sensitivity

Active Publication Date: 2021-01-05
JINAN UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, only a small number of patients treated with imatinib achieved complete remission, the remaining 20% ​​of patients discontinued the drug due to drug intolerance, and another 20% developed drug resistance that limited its use
Although, the newly marketed TKIs in recent years, such as nilotinib, dasatinib, bosutinib and ponatinib, provide a variety of options for CML patients, and these new drugs show stronger Efficacy, but still failed to solve the problem of drug resistance in patients

Method used

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  • SiRNA for targeted inhibition of SOS1 gene expression and application of siRNA
  • SiRNA for targeted inhibition of SOS1 gene expression and application of siRNA
  • SiRNA for targeted inhibition of SOS1 gene expression and application of siRNA

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Experimental program
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Embodiment 1

[0025]1. Materials and methods

[0026]1. Material

[0027]1.1 Cells and animals

[0028]K562, KCL-22 (human chronic myeloid leukemia), purchased from Shanghai Cell Bank of Chinese Academy of Sciences, BV173 (human peripheral blood B-cell leukemia), donated to the American Institute of Hematology, and can also be obtained from commercial sources. KU812 (Human Chronic Myeloid Leukemia), donated to the Pan Jingxuan Research Group of Jinan University, can also be obtained from commercial sources. Balb / cnude mice aged 4-6 weeks were purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.

[0029]1.2 Reagents

[0030]1640 medium, opti-MEM medium, anti-SOS1 (mouse SOS1 monoclonal antibody) (santa), anti-β-actin (rabbit β-actin monoclonal antibody) (CST), siRNA freeze-dried powder (sharp BioBio), CCK-8 (biomake), imatinib (STI571) (selleck), lipo2000 (thermo), qPCR reagent (biomake).

[0031]1.3 Random sequence of RNA duplex and SOS1-siRNA sequence

[0032]1.3.1 Random sequence of double-st...

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Abstract

The invention discloses siRNA for targeted inhibition of SOS1 gene expression and application of the siRNA. The sequence of the siRNA is as follows: a sense strand is 5'-GCAGAATCTTCACCATCTA-3', and anantisense strand is 5'-TAGATGGTGAAGATTCTGC-3'. According to the SiRNA, by detecting the change of expression quantity of the mRNA level and protein level of SOS1, the effective siRNA sequence of SOS1, namely the SOS1-siRNA # 3 is screened out, the multiplication capacity and clonality of CML cells can be effectively inhibited, the growth of transplanted tumors can be effectively inhibited in animal bodies, and meanwhile, the drug sensitivity of the CML cells to imatinib is improved. Therefore, the SOS1 is a potential new target for treating chronic granulocytic leukemia, and the SOS1 siRNA has a potential value of overcoming the drug resistance of the imatinib.

Description

Technical field[0001]The invention relates to a siRNA, in particular to a siRNA targeted to inhibit SOS1 gene expression and its application.Background technique[0002]Chronic myelogenous leukemia (CML) is a characteristic genetic abnormality, and the ph chromosome is its characteristic cytogenetic marker, namely t(9;22)(q34;ql1), and there is a specific BCR-ABL fusion gene. A disease that causes malignant clonal proliferation of hematopoietic stem cells. Imatinib, as the first generation of tyrosine kinase inhibitor that targets the P210 protein encoded by the BCR-ABL fusion gene, has revolutionized the treatment of chronic granulocytes. However, only a small number of patients receiving imatinib have achieved complete remission, the remaining 20% ​​of patients stopped taking the drug due to drug intolerance, and 20% of patients developed drug resistance and restricted their use. Although, in recent years, newly marketed TKIs, such as nilotinib, dasatinib, bosutinib, and panatinib, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61K31/713A61P35/02
CPCC12N15/113A61K31/713A61P35/02C12N2310/141
Inventor 费嘉刘燕君
Owner JINAN UNIVERSITY
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