Application and method of USP10 gene and/or Ascl1 gene in inducing transdifferentiation of fibroblasts into neuronal cells

A technology of fibroblasts and neuron cells, applied in the biological field, can solve problems such as the inability to obtain neuron cells efficiently, and achieve the effect of improving stability

Pending Publication Date: 2022-03-25
INST OF ZOOLOGY CHINESE ACAD OF SCI
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the deubiquitination process of Ascl1 has not been reported
[0006] Therefore, it is of great significance for the treatment of nerve-related diseases to devel

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application and method of USP10 gene and/or Ascl1 gene in inducing transdifferentiation of fibroblasts into neuronal cells
  • Application and method of USP10 gene and/or Ascl1 gene in inducing transdifferentiation of fibroblasts into neuronal cells
  • Application and method of USP10 gene and/or Ascl1 gene in inducing transdifferentiation of fibroblasts into neuronal cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Screening to improve the stability of Ascl1 protein and USP10 to Ascl1 deubiquitination

[0041] 1. Stability detection of Ascl1 protein

[0042] Overexpressed in mouse embryonic fibroblasts (MEFs), human embryonic kidney cells (293T), mouse neuroblastoma cells (N2A) and mouse neural stem cells (NSCs) by lipofection or viral infection Ascl1, the specific steps are as follows:

[0043] Construct the overexpression plasmid of Ascl1 (comprising PCDH-Ascl1 plasmid and pAD-Ascl1 plasmid): The construction of PCDH-Ascl1 plasmid is as follows: first, the Ascl1 gene fragment is amplified in the cDNA library of mouse embryonic period (E13) brain, amplified The primers are: Ascl1-Xba1-F: 5'-TCTAGAATGGAGAGCTCTGGCAAGATG-3' (as shown in SEQ ID NO.3); Ascl1-Ecor1-R: 5'-GAATTC TCAGAACCAGTTGGTAAAGTCC-3' (as shown in SEQ ID NO.4) ; Then use restriction endonucleases Xba1 and EcoR1 to digest the pCDH plasmid and the Ascl1 gene fragment respectively. The digested PCDH plasmid...

Embodiment 2

[0057] Example 2 Inducing Fibroblasts to Transdifferentiate into Neuronal Cells

[0058] 1. Adenoviral vector construction

[0059] First, the USP10 gene (CDS sequence, as shown in SEQ ID NO.1) and the Ascl1 gene (CDS sequence, as shown in SEQ ID NO.2) were cut by restriction endonucleases Ecor1 and Kpn1 and connected by T4 ligase. Connect to pENTR TM 3C-GFP Dual Selection Vector (pENTR) (Invitrogen) vector to obtain pENTR-USP10 and pENTR-Ascl1 plasmids. Then use pENTR TM 3C-GFP Dual SelectionVector and the recombination sequence on the pAD vector, construct the gene sequence of USP10 and Ascl1 on the adenovirus pAD vector by means of recombinase (CRE) recombination, that is, by using the LRCionase II enzyme mixture to make the gene sequence on the pAD vector LR homologous recombination occurs between attR1 and attR2 and attL1 and attL2 on the PEIG plasmid, and the exogenous gene is transferred to the pAD vector. The reaction system is 1 μL pAD vector, 1 μL pENTR-USP10 or...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses an application and a method of a USP10 gene and/or an Ascl1 gene in inducing transdifferentiation of fibroblasts into neuronal cells. The invention firstly discloses the application of the USP10 gene and/or the Ascl1 gene in inducing the fibroblast to be transdifferentiated into the neuronal cell or the application in preparing a product for inducing the fibroblast to be transdifferentiated into the neuronal cell. The invention further discloses a method for inducing transdifferentiation of fibroblasts into neuronal cells. The USP10 capable of improving the stability of the Ascl1 protein and deubiquitinating the Ascl1 is obtained by screening based on the characteristics of the Ascl1. The USP10 gene and the Ascl1 gene are applied to the process of converting fibroblasts into neuronal cells, the GABAergic neuronal cells can be efficiently obtained, the defect that the neuronal cells cannot be efficiently obtained is overcome, and the application has important significance on treatment of nerve-related diseases.

Description

technical field [0001] The present invention relates to the field of biotechnology. More specifically, it relates to the application and method of USP10 gene and / or Ascl1 gene in inducing the transdifferentiation of fibroblasts into neuron cells. Background technique [0002] Human beings and most animals and plants are composed of many cells. These cells gradually form specific shapes and functions during development, and finally achieve precise functions in each organ. These cells are called differentiated cells. Ordinary cells retain their specificity until they die, but under certain circumstances, some cells can change state to replace another specific cell to function, which is called "transdifferentiation". If an effective method can be found to transform differentiated functional adult cells into another functional cells, it will be of great significance for the treatment of many major diseases. [0003] In February 2010, Vierbuchen et al. (Vierbuchen T, Ostermeier...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N5/10C12N5/0793C12N15/861C12N15/57C12N15/12
CPCC12N5/0619C12N5/0625C12N15/86C12N9/485C07K14/4702C12Y304/19012C12N2710/10043C12N2506/09C12N2510/00C12N2501/13C12N2500/38C12N2501/01Y02A50/30
Inventor 焦建伟张东明
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap