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Use of human ckip1 gene and related medicines

A gene and application technology, applied in the application of human CKIP1 gene and its related medicines, can solve the problem that PLEKHO1 function has not been reported yet

Active Publication Date: 2020-09-22
SHANGHAI GENECHEM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research on the correlation between PLEKHO1 and cancer has just begun, and there is no report on the function of PLEKHO1 in the proliferation and malignant progression of liver cancer, gastric cancer, breast cancer, breast cancer and glioma

Method used

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  • Use of human ckip1 gene and related medicines
  • Use of human ckip1 gene and related medicines
  • Use of human ckip1 gene and related medicines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Example 1 Preparation of RNAi lentivirus against human CKIP1 gene

[0087] 1. Screening for effective siRNA targets against human CKIP1 gene

[0088] Retrieve CKIP1 (NM_016274.4) gene information from Genbank; design effective siRNA targets for CKIP1 gene. Table 1 lists 17 effective siRNA target sequences for CKIP1 gene.

[0089] Table 1 is targeted at the siRNA target sequence of human CKIP1 gene

[0090] SEQ ID NO. TargetSeq start site 1 GCTGAGAGACCTGTACAGA 1411 2 GGACCTGGTAGCAAGGAAA 1204 3 GTGACTATGAGAAGTGTGA 495 4 TTACTCTTTGCCCACTCCAA 566 5 AAGTTTACTTCTTGCCCACT 562 6 TTACTCTTTGCCCACTCCAA 560 7 CAAGAACCGTATCTTGGAT 684 8 GAGGCATCATCGAATTGGA 1363 9 GAAGGAATCGTGGATCAAT 639 10 CCAGAAGAGAAGGAATCGT 631 11 TGCTGACCTTGGACTTGAT 833 12 GAGACCTGTACAGACAGAT 1421 13 ATTTGACCTGAGTGACTAT 489 14 CAAGAACCGTATCTTGGAT 678 15 GAGGCATCATCGAATTGGA 1357 16 GAGTCCAGA...

Embodiment 2

[0112] Example 2 Real-time fluorescent quantitative RT-PCR method to detect the silencing efficiency of CKIP1 gene

[0113] Human lung cancer H1299 cells, glioma U87 cells and liver cancer SMMC-7721 cells in the logarithmic growth phase were trypsinized to make cell suspension (the number of cells was about 5×10 4 / ml) were seeded in 6-well plates and cultured until the cell confluency reached about 30%. According to the multiplicity of infection (MOI, H1299:1; U87:2; SMMC7721:10) value, an appropriate amount of the virus prepared in Example 1 was added, and the culture medium was replaced after 24 hours of incubation. After the infection time reached 5 days, the cells were collected. Total RNA was extracted according to the instruction manual of Invitrogen's Trizol. According to Promega's M-MLV operating instructions, RNA was reverse-transcribed to obtain cDNA (see Table 7 for the reverse transcription reaction system, react at 42 °C for 1 h, and then bathe in a water bath a...

Embodiment 3

[0120] Example 3 Detection of proliferation ability of tumor cells infected with CKIP1-siRNA lentivirus

[0121] Human lung cancer H1299 cells, glioma U87 cells and liver cancer SMMC-7721 cells in the logarithmic growth phase were trypsinized to make cell suspension (the number of cells was about 5×10 4 / ml) were seeded in 6-well plates and cultured until the cell confluency reached about 30%. According to the multiplicity of infection (MOI, H1299:1; U87:2; SMMC7721:10), add an appropriate amount of virus, replace the medium after 24 hours of culture, and collect the experiments in the logarithmic growth phase after the infection time reaches 5 days group of cells. The complete medium was resuspended into a cell suspension (2×10 4 / ml), inoculate a 96-well plate at a cell density of about 2000 cells / well. 5 replicate wells in each group, 100 μl per well. After laying the board, place at 37°C, 5% CO 2 Incubator cultivation. From the second day after plating, the plate was...

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Abstract

The invention discloses a use and related drugs of a human CKIP1 gene. The invention discloses the use of the human CKIP1 gene in tumor therapy, tumor diagnosis and drug preparation. The invention further builds a human CKIP1 gene small-molecular interfering RNA, a human CKIP1 gene interfering nucleic acid building body and a human CKIP1 gene interfering lentivirus and uses thereof. The siRNA or the siRNA sequence-containing nucleic acid building body and lentivirus can specifically inhibit expression of the human CKIP1 gene, and especially the lentivirus can efficiently infect target cells, efficiently inhibit expression of the CKIP1 gene in the target cells, thereby inhibiting the growth of tumor cells, promoting tumor cell apoptosis, and having important significance in tumor therapy.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to the use of human CKIP1 gene and related medicines. Background technique [0002] The phenomenon of ribonucleic acid interference (RNA interference, RNAi) refers to the specific degradation of the mRNA when a double-stranded RNA (dsRNA) homologous to a certain sequence of the endogenous mRNA coding region is introduced into the cell, resulting in the silencing of the gene expression . Studies have shown that double-stranded RNA with a length of 21-23nt can specifically cause RNAi at the transcriptional and post-transcriptional levels (TuschlT, Zamore PD, Sharp PA, Bartel DP.RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at21to23nucleotide intervals. Cell 2000;101:25-33.). Tumor is a major disease that threatens human health. Although cancer patients have undergone chemotherapy, radiotherapy and comprehensive treatment, their five-year survival ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/867A61K31/713A61P35/00
CPCA61K48/0016C12N15/1135C12N15/867C12Q1/6886
Inventor 朱向莹孙琴谭畅吴涛金杨晟瞿红花曹跃琼
Owner SHANGHAI GENECHEM
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