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Lenvatinib drug-resistant gene NF1 as well as screening method and application thereof

A technology of lenvatinib and drug-resistant genes, which is applied in the fields of application, genetic engineering, and plant genetic improvement, and can solve problems such as off-target effects, unstable and obvious phenotypic changes, etc.

Pending Publication Date: 2021-06-22
柳州市柳铁中心医院
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Problems solved by technology

However, the phenotypic changes caused by RNAi technology are not stable and obvious, and the endogenous RNAi pathway of cells leads to a wide range of off-target effects in RNAi screening

Method used

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  • Lenvatinib drug-resistant gene NF1 as well as screening method and application thereof
  • Lenvatinib drug-resistant gene NF1 as well as screening method and application thereof
  • Lenvatinib drug-resistant gene NF1 as well as screening method and application thereof

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Embodiment Construction

[0060] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0061] The lenvatinib resistance gene of the embodiment of the present invention is NF1, and the sequence of the gene is SEQ ID NO:1.

[0062] see Figure 1 to Figure 21 , the screening method of lenvatinib drug-resistant gene NF1 of the present invention comprises:

[0063] Using the HuH7 cell line as the target cell, the high-throughput functional screening technology of CRISPR-Cas9 was used to screen the differential target genes of lenvatinib resistance. The steps are as follows:

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Abstract

The invention relates to the technical field of tumor drug-resistant genes, in particular to a lenvatinib drug-resistant gene NF1 as well as a screening method and application thereof. The gene sequence of the NF1 is SEQ ID NO: 1. The screening method comprises the following steps: carrying out a pre-experiment to obtain the optimal MOI of the sgRNA library, and determining the concentration of lenvatinib; carrying out an infection Cas9 library experiment to obtain a stable strain; adding lenvatinib into the stable strain for treatment, and analyzing the enrichment condition of the sgRNA through PCR amplification and high-throughput sequencing; screening a drug-resistant gene of lenvatinib; and verifying the function of the effective target gene on liver cancer cell function phenotype and mediated lenvatinib drug resistance. The drug-resistant gene of the lenvatinib is obtained through CRISPR / Cas9 whole-genome library screening, a theoretical basis is provided for reducing the drug resistance of the lenvatinib in the future, a new drug-resistant target is provided for clinical application of the lenvatinib, and guidance is provided for clinical reasonable medication.

Description

technical field [0001] The invention relates to the technical field of tumor drug resistance genes, in particular to a lenvatinib drug resistance gene NF1, its screening method and application. Background technique [0002] Liver cancer is one of the most common malignant tumors, its incidence rate ranks third among all tumors in my country, and its mortality rate ranks second. On September 4, 2018, Lenvatinib was launched in China as a first-line treatment for advanced liver cancer, and it was officially replaced by Lenvatinib as a first-line treatment for advanced liver cancer Sorafenib has become the drug of choice for the first-line treatment of advanced liver cancer in the world. It is a small molecule multi-target tyrosine kinase inhibitor. Small-molecule multi-target tyrosine kinase inhibitors have anti-tumor cell proliferation and anti-angiogenesis effects. The main mechanisms of targeted drug resistance include the establishment of compensatory signaling pathways i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/867C12N15/10C12Q1/02
CPCC07K14/47C12N15/1082C12N15/86G01N33/5011C12N2740/15043C12N2800/107
Inventor 孙一帆陈坤沈永奇卢永刚张洁何沙孙林黄文杰肖笑荣
Owner 柳州市柳铁中心医院
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