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Construction method and application of Ifit3-eKO1 gene knockout mouse animal model

A gene knockout mouse, ifit3-eko1 technology, applied in the biological field, can solve the problem that the construction method has not been reported

Active Publication Date: 2017-12-15
SHANGHAI TONGJI HOSPITAL
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Problems solved by technology

However, there is no report on the construction method of Ifit3-eKO1 gene knockout mouse animal model

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  • Construction method and application of Ifit3-eKO1 gene knockout mouse animal model
  • Construction method and application of Ifit3-eKO1 gene knockout mouse animal model
  • Construction method and application of Ifit3-eKO1 gene knockout mouse animal model

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Embodiment Construction

[0047] The specific embodiments provided by the present invention will be described in detail below in conjunction with the accompanying drawings.

[0048] 1. This implementation category involves a method for establishing an Ifit3-eKO1 gene knockout mouse model based on CRISPR / Cas9 gene knockout technology. The technical route is as follows: figure 1 shown.

[0049] 2. The basic information of knockout gene

[0050] 1. Knockout gene name (MGI number): Ifit3 (1101055)

[0051] 2. Knockout gene MGI website link: http: / / www.informatics.jax.org / marker / MGI:1101055

[0052] 3. Knockout gene name (Ensembl): Ifit3 (ENSMUSG00000074896)

[0053]4. Knockout gene Ensembl website link: http: / / asia.ensembl.org / Mus_musculus / Gene / Summary? g=ENSMUSG00000074896; r=19:34583531-34588731; t=ENSMUST00000102825

[0054] 5. Knockout the targeted transcript (Ensembl number): Ifit3-001 (ENSMUST00000102825.3)

[0055] Knockout targeted exon: exon 2

[0056] 3. Schematic diagram of the design str...

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Abstract

The invention relates to a method of constructing an Ifit3-eKO1 gene knockout mouse animal model and belongs to the biotechnical field. The method comprises the following steps: S1, determining specific target sites sgRNA1 and sgRNA2 of a to-be-knocked out gene of an Ifit3-eKO1 mouse, and performing in vitro transcription with Cas9 nuclease to mRNA; and S2, micro-injecting active sgRNA and Cas9RNA into an oosperm of the mouse to obtain the Ifit3-eKO1 gene knockout mouse. The method has the advantages that by using the CRISPR / Cas9 gene knockout technology, the Ifit3-eKO1 gene knockout mouse animal model is constructed for the first time, thereby providing a convenient, reliable and economical animal model for researching action of Ifit3 in tumorigenesis and development.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for constructing an Ifit3-eKO1 gene knockout mouse animal model and its application. Background technique [0002] The Ifit3 gene, also known as the RIG-G gene (retinoic acid-induced gene G, retinoic acid.induced geneG), is cloned from the acute promyelocytic leukemia (acute promyelocytic leukemia, APL) cell line NB4 and can be fully The gene induced by trans retinoic acid (all-trans retinoic acid, ATRA) is located on chromosome 10 q24, encoding a protein with 490 amino acid residues and a molecular weight of about 60,000. [0003] Although Ifit3 was originally cloned from the acute promyelocytic leukemia cell line NB4, bioinformatic analysis revealed that Ifit3 is a member of the interferon-induced gene family. It has been reported that Ifit3 can not only be significantly induced by ATRA in NB4 cells, but also can be rapidly induced by interferon in a variety of tumor cell...

Claims

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Application Information

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IPC IPC(8): C12N15/90A01K67/027C12N15/113
CPCA01K67/0276A01K2217/075A01K2227/105A01K2267/03C07K14/47C12N15/113C12N15/907C12N2310/10
Inventor 李冬吴军录权文强
Owner SHANGHAI TONGJI HOSPITAL
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