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sgRNA and gene vector for inhibiting bladder cancer by targeting human lncRNA-UCA1 and application of sgRNA

A gene carrier, the technology of pgl3-u6-uca1sgl, is applied to the sgRNA targeting human lncRNA-UCA1 to inhibit bladder cancer, the gene carrier and its application field, which can solve the problems of easy drug resistance, easy recurrence of patients, obvious side effects, etc. , to achieve the effect of inhibiting tumor growth, high knockout efficiency and simple steps

Active Publication Date: 2017-02-15
THE FIRST AFFILIATED HOSPITAL OF XIAN JIAOTONG UNIV
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Problems solved by technology

[0006] The existing problems in the treatment of human bladder cancer: (1) patients often relapse after treatment; (2) drug resistance is easy to develop after drug treatment; (3) there are few therapeutic drugs targeting human bladder cancer oncogenes, and the effect is limited. The side effects are obvious; (4) combination therapy is needed to treat bladder cancer to improve the therapeutic effect and improve the patient's prognosis

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  • sgRNA and gene vector for inhibiting bladder cancer by targeting human lncRNA-UCA1 and application of sgRNA

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Embodiment Construction

[0032] The present invention will be described in detail below in conjunction with the accompanying drawings and embodiments.

[0033] like figure 1 As shown, the directional recognition and cleavage of genes by the CRISPR / Cas9 system are realized by sgRNA and Cas9, and sgRNA determines the targeting of Cas9 and also determines the cleavage activity of Cas9. The present invention aims to apply CRISPR / Cas9 technology, take human bladder cancer cells with high expression of lncRNA-UCA1 and bladder cancer xenograft mouse model as research objects, use target pathogenic gene lncRNA-UCA1, and coordinate tumor immune regulation Gene editing strategies for the molecule PD-1. Firstly, the lncRNA-UCA1 gene is effectively knocked out by screening the sgRNA sequence targeting the lncRNA-UCA1 gene in vivo and in vitro; then, the sgRNA sequence targeting PD-1 is screened in vivo and in vitro, and the multi-gene cooperative editing effect is realized by co-transfection. Furthermore, it wa...

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Abstract

The invention provides sgRNA and a gene vector for inhibiting bladder cancer by targeting human lncRNA-UCA1 and an application of the sgRNA; specifically, sgRNA sequences of an lncRNA-UCA1 gene and a PD-1 gene suitable for CRISPR-Cas9 targeted splicing are designed; by transforming plasmids of specific splicing lncRNA-UCA1 gene and CRISPR-Cas9 nuclease gene into human bladder cancer cells, the expression of the lncRNA-UCA1 gene drops so as to inhibit the growth of tumor cells; and the plasmid, together with a human PD-1 gene targeted knockout vector, is transformed into a humanized mouse model of bladder cancer transplanted tumor, so as to obviously inhibit the growth of tumors. According to the invention, the vector is simple in preparing steps, the sgRNA is good in targeting property and the CRISPR-Cas9 is high in knockout efficiency.

Description

technical field [0001] The invention belongs to the field of genetic engineering and biomedicine, and relates to the innovative design of CRISPR / Cas9 specific modification of human long-chain non-coding RNA (lncRNA)-UCA1 and multiple gene targets of PD-1, through targeted knockout of the expression of lncRNA UCA1 Inhibiting the growth of bladder cancer cells, combined with the application of immune gene (PD-1 / PDL-1) therapy strategy can enhance the therapeutic effect of bladder cancer, which is a new strategy of targeting immune gene therapy for bladder cancer. Background technique [0002] In recent years, genome editing tools have been widely used in the field of biomedicine, which can change the expression of target genes, elucidate the function of genes, and try to treat clinical diseases. Among them, clustered regularly interspaced short palindromic repeats (CRISPR) technology can quickly, easily and efficiently target any gene in the genome, and has the advantages of e...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/85A61K31/7088A61K48/00A61P35/00
CPCC07K14/70521C07K14/82C12N15/1135C12N15/1138C12N15/85C12N2310/10C12N2800/107C12N2800/80C12N2810/10
Inventor 李旭陈葳甄帅赵乐
Owner THE FIRST AFFILIATED HOSPITAL OF XIAN JIAOTONG UNIV
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