PROM1-KO mouse model construction method and application thereof

A mouse model and construction method technology, applied in the fields of molecular biology and biomedicine, can solve the problem of not fully showing pathological manifestations, and achieve the effect of high knockout efficiency, reliable theoretical basis, and no foreign gene interference.

Active Publication Date: 2019-09-20
上海朗昇生物科技有限公司
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  • Abstract
  • Description
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Problems solved by technology

Defect: The imported PROM1 is of human origin. Due to certain differences between human and mouse PROM1 ge...

Method used

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  • PROM1-KO mouse model construction method and application thereof
  • PROM1-KO mouse model construction method and application thereof
  • PROM1-KO mouse model construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Construction and Identification of PROM1-KO Mouse Model

[0055] 1. Materials

[0056] The mouse strain used in this example is: C57BL / 6J, and the surrogate mother mouse strain is C57BL / 6J, which was purchased from Shanghai Slack Experimental Animal Co., Ltd., and the mice were randomly divided into a control group and an experimental group.

[0057] 2. Method

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Abstract

The invention relates to a PROM1-KO mouse model construction method and application thereof. A mouse model is a PROM1 gene knockout mouse. A PROM1 gene knockout mouse model is constructed based on CRISPR/Cas9 gene knockout technology. The construction method comprises the following steps: step one, designing sgRNA and Cas9mRNA, transcribing sgRNA and Cas9mRNA into mRNA in vitro, and microinjecting the active sgRNA and the active Cas9mRNA into mouse fertilized eggs to obtain PROM1-KO mice; and step two, identifying a PROM1-KO mouse animal model. The PROM1-KO mouse animal model is firstly constructed based on the CRISPR/Cas9 gene knockout technology, and a convenient, reliable and economical animal model is provided for researching the relationship between PROM1 and diseases such as hereditary retinopathy and tumors.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and biomedicine, in particular to a method for constructing a PROM1-KO mouse model and its application. Background technique [0002] The CRISPR / Cas system is developed from the adaptive immune system of bacteria and archaea against foreign viruses or plasmids, including three different types, of which the Type II CRISPR / Cas system has only one subunit of the DNA endonuclease Cas9 , the structure is the simplest, so it is the most widely used. In addition to the Cas9 protein, the system also includes two short CRISPR RNAs (crRNAs) and trans-activatingcr RNAs (tracrRNA). The mature crRNA-tracrRNA complex can guide the Cas9 protein to the target sequence through complementary base pairing, and specifically cut the DNA double strand near the PAM (protosp aceradjacent motif) to form a DSB (double strand break). DSB can be repaired in two ways, one is Non-Homologous End Joining (NHEJ) DNA ...

Claims

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Application Information

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IPC IPC(8): C12N15/90C12N9/22A01K67/027C12N5/10C12N15/113
CPCA01K67/0276A01K2217/075A01K2227/105A01K2267/0306A01K2267/0331C07K14/70596C12N9/22C12N15/1138C12N15/907C12N2310/20
Inventor 梁坚
Owner 上海朗昇生物科技有限公司
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