Hairless model pig reconstructed oocyte and reconstruction method thereof and model pig construction method

A construction method and model technology, applied in the field of animal model construction, can solve the problems of difficulty in somatic cell targeting, low efficiency of gene-directed modification in large animals, etc., and achieve the effect of avoiding phenotypic differences

CN106957856AActive Publication Date: 2017-07-18广州中科飞豚生物科技有限责任公司
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2017-07-18

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Abstract

The invention relates to mutation site controllable hairless model pig reconstructed oocyte and a reconstruction method thereof and a model pig construction method. According to the hairless model pig reconstructed oocyte and reconstruction method thereof and the model pig construction method, by utilizing CRISPR / Cas9 and point mutation technology, cells can be accurately repaired by utilizing provided ssODNs as a template in a homologous recombination manner after Cas9 is cut at the gene specific site, and a termination codon is introduced at the gene specific site through the CRISPR / Cas9 and ssODNs design, so that gene translation is early terminated, and further the loss of gene function can be caused. Human disease gene mutation type can be accurately replicated by utilizing the method, so that the phenotype difference caused due to different gene mutation forms can be avoided. A hairless model pig constructed by utilizing the method has no other abnormities except for hair loss, hoof development defect and filiform papillae development defect, and can be used for various dermatological researches and external skin drugs and cosmetics. The skin transformed immunologically can be used for clinical skin transplantation.
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Description

technical field

[0001] The invention relates to the field of animal model construction, in particular to a reconstituted egg of a hairless model pig, a construction method thereof, and a construction method of a model pig. Background technique

[0002] Biomedical research cannot do without animal models. Unlike humans, other mammals are mostly covered with thick hair, which is not conducive to dermatological research. Establishing a hairless animal model, in addition to avoiding skin damage caused by hair removal during the experiment, is mainly used for skin transplantation, skin aging and anti-aging research, cosmetics and external skin drug testing. In addition, hairless animals can be used as research models for alopecia, low-onset diseases. According to investigations, the incidence rate of male hair loss in my country is as high as 25%. With the acceleration of the pace of life, the enhancement of work pressure, and the change of diet and living habits, the incidenc...

Examples

Embodiment Construction

[0042] In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the associated drawings. Preferred embodiments of the invention are shown in the accompanying drawings. However, the present invention can be embodied in many different forms and is not limited to the embodiments described herein. On the contrary, these embodiments are provided to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0043] In this embodiment, it is found through experiments that the PHNED-related gene HOXC13 is knocked out using Crispr / Cas9 technology and point mutation technology to establish a hairless model pig with pure hair-nail ectodermal dysplasias (PHNED) . The hairless cloned pig model established in this embodiment has no other developmental abnormalities except for the disappearance of body hair and abnormal development of hooves and tongue papillae. Th...