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Composition and method for inducing somatic cells to transdifferentiate into sensory neurons and ganglions

A transdifferentiation, neuron technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, embryonic cells, etc., can solve the problem of unreported sensory neuron types and so on

Active Publication Date: 2019-11-19
向孟清 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Pain sensory neurons have been differentiated from human PSCs, and these neurons can respond to the stimulation of P2XR3 and capsaicin [2], but no other differentiated sensory neuron types have been reported

Method used

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  • Composition and method for inducing somatic cells to transdifferentiate into sensory neurons and ganglions
  • Composition and method for inducing somatic cells to transdifferentiate into sensory neurons and ganglions
  • Composition and method for inducing somatic cells to transdifferentiate into sensory neurons and ganglions

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Embodiment Construction

[0057] The specific process and details of the present invention will be described in detail below by taking "Transdifferentiation of mouse fibroblasts into sensory neurons by Ascl1 and Isl1 lentiviruses" as an example.

[0058] (1) Construction of lentiviral vector (plasmid)

[0059] The mouse Ascl1 (NM_008553.4, 696bp) and Isl1 (NM_021459.4, 1050bp) coding sequences were cloned into different lentiviral vectors (Tet-O-FUW) containing the tet-on regulatory sequence to obtain the plasmid Tet-O - FUW-Ascl1, and plasmid Tet-O-FUW-Iscl1. The purpose is to co-transfect with RTTA lentivirus in the future, and the drug Dox (Doxycycline) can be added to control gene expression in somatic cells. After the correct cloning was verified by DNA sequencing, the plasmids for transfection were prepared.

[0060] (2) Preparation of lentivirus

[0061] 1) 24 hours before plasmid transfection, spread 4.0x10 cells in a 10cm cell culture dish 6 HEK293T cells were cultured by adding 6ml MEF me...

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Abstract

The invention discloses a composition and method for inducing somatic cells to transdifferentiate into sensory neurons and ganglion-like structures. The composition includes any combined transcriptionfactor of Ascl1+Isl1, Ascl1+Brn3a, Ascl1+Brn3b, Ascl1+Brn3c, Ascl1+Isl1+Brn3a, Ascl1+Isl1+Brn3b and Ascl1+Isl1+Brn3c, can induce the somatic cells to transdifferentiate into the sensory neurons and the ganglion-like structures.

Description

technical field [0001] The present invention relates to the fields of biotechnology such as cell fate transdifferentiation, sensory neuron disease research, etc. It mainly uses transcription factors to induce transdifferentiation of human or animal somatic cells into central / peripheral sensory neurons and ganglion-like structures in vitro, providing A new way to obtain sensory neurons induced in vitro. Background technique [0002] Sensory neurons can sense external signals and can be divided into vision, auditory, taste, smell, temperature, and mechanical stimulation (mechanoreceptors), and can also sense internal signals including blood Receptors (blood receptors), nociceptors (nociceptors), etc. Nociceptors are divided into heat and cold receptors and other subtypes. According to the location distribution, sensory neurons can be divided into central and peripheral sensory neurons. Central sensory neurons include sensory neurons that perceive vision, hearing, smell, and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079C12N5/0793C12N5/073C12N15/867C12N5/10
CPCC12N5/0618C12N5/0619C12N15/86C12N2501/60C12N2506/02C12N2506/1307C12N2740/15043C12N2800/107C12N2510/00Y02A50/30
Inventor 向孟清晋康新邹珉肖冬长
Owner 向孟清
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