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Melanoma cell strain and preparation method thereof

A cell line and melanoma technology, applied in the field of melanoma cell line and its preparation, can solve the problems of melanoma gene instability, affecting the research process, long reproduction cycle, etc., and achieve the effect of stabilizing the genetic function

Pending Publication Date: 2022-04-05
CHINA JAPAN FRIENDSHIP HOSPITAL
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  • Application Information

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Problems solved by technology

However, the existing MM transgenic mice have a long reproductive cycle, and the melanoma gene in the body is not stable after breeding, which affects the progress of the research

Method used

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  • Melanoma cell strain and preparation method thereof
  • Melanoma cell strain and preparation method thereof
  • Melanoma cell strain and preparation method thereof

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Embodiment Construction

[0023] The present invention will be further described in detail below in conjunction with the embodiments, so that those skilled in the art can implement it with reference to the description.

[0024] 1. Contains Rosa26-Stop-tdTomato, Tyr-creER T2 、Braf V600E and Pten f / f Preparation of genetic mice

[0025] First, FVB / NJ mice were selected, and the fusion gene was injected into the fertilized egg cells of the mice by transgenic microinjection technology to construct Tyr-creER-containing T2 Genetic mice, that is, to construct a melanocyte-specific cre enzyme active mouse, so that Tyr-creER T2 Genetic mice with Braf that can be conditionally driven in vivo V600E Mutation Activation and Pten Gene Loss (Pten f / f ) mouse hybridization, that is, to obtain Tyr-creER containing T2 、Braf V600E 、Pten f / f Genetic mice that will contain Tyr-creER T2 、Braf V600E 、Pten f / f Gene mice are crossed with mice containing the Rosa26-Stop-tdTomato gene, and Rosa26-Stop-tdTomato, Tyr-cr...

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Abstract

The invention discloses a melanoma cell strain, the melanoma cell strain contains a Rosa26-Stop-tdTomato reporter gene, a Tyr-creERT2 gene, a BrafV600E and a Ptenf / f gene, the melanoma cell strain is classified and named as a mouse melanoma cell strain ATBP, the cell strain is preserved in the China Center for Type Culture Collection on February 9, 2022, and the preservation number is CCTCC NO: C202233. The invention also provides a preparation method of the melanoma cell strain. The melanoma cell strain which is stable in genetic function, capable of expressing red fluorescent protein and contains the BrafV600E mutation activation gene and the Pten deletion gene can be prepared, and medical research is facilitated.

Description

technical field [0001] The invention relates to the field of biotechnology, more specifically, to a melanoma cell line and a preparation method thereof. Background technique [0002] Cutaneous malignant melanoma (MM for short) is the deadliest malignant tumor in dermatology, which is formed by malignant transformation of melanocytes located in the basal layer of the epidermis. The generation of MM is caused by the interaction of genetic factors and environmental factors. Generally, the genes that are prone to mutations in melanocytes to form MM are BRAF, NRAS, TP53, PTEN, CDKN2A, and KIT. Among them, BRAF mutations in melanocytes The probability is 63%, and the probability of PTEN deletion is 25%. [0003] In order to study the development process of MM, evaluate the effect of tumor drug treatment and the mechanism of drug resistance, etc., MM transgenic mice are often used for research. However, the breeding cycle of the existing MM transgenic mice is long, and the melano...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/12C12N15/55A01K67/027C12R1/91
Inventor 许静凯刘宝怡李昂左先波崔勇
Owner CHINA JAPAN FRIENDSHIP HOSPITAL
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