Method for knocking out NANS gene in iPSC (induced pluripotent stem cells) and application

A cell and gene technology, applied in the field of cell model construction, can solve problems such as brain organoid mental development disorder models that have not yet been seen, and achieve the effect of improving gene editing efficiency, complete and thorough effect, and thorough knockout effect
CN111662907AActive Publication Date: 2020-09-15成都华西海圻医药科技有限公司 +1
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
成都华西海圻医药科技有限公司
Publication Date
2020-09-15

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Abstract

The invention discloses a method for knocking out an NANS gene in iPSC (induced pluripotent stem cells) and application, and belongs to the field of cell model construction. According to the invention, a CRIPSR-Cas9 gene editing system is transferred into the iPSC by adopting an electroporation method; Donor DNA containing a Puromycin resistance gene and a GFP gene is inserted into an NANS exon, and the stem cells subjected to gene editing are screened by Puromycin; and after transfection, CloneRTM culture cells are added, and the system can greatly improve gene editing efficiency of a CRISPR-Cas9 genome editing technology in the stem cells. The method is helpful for constructing an NANS defect type 3D brain organ model, and can be applied to in-vitro study and drug research and development on the basis of pathogenesis of mental development disorders caused by the NANS mutation.
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Description

technical field

[0001] The invention relates to the field of cell model construction, in particular to a method and application for knocking out the NANS gene of induced pluripotent stem cells based on CRISPR-CAS9 gene editing technology. Background technique

[0002] Intellectual developmental disorders (Intellectual developmental disorders, IDD), also known as mental retardation, mental retardation C Mental Retardation, MR), is a group of onset before the age of 18, cognitive dysfunction (IQ value <70 points), social A disorder characterized by deficits in adaptive capacity. In preschool age (below 5 years old), it is manifested as language ability and motor development delay, which is also known as developmental delay (Developmental Delay, DD). Syndromic mental retardation is usually accompanied by multiple congenital anomalies (Multiple congenital anomalies, MCA), special face and so on. The etiology of ID / DD is complex, including environmental factors, perinatal hy...

Claims

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