RE1-Binding Expression Vector for In Vivo Glial Reprogramming

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Existing methods for inducing neuronal transdifferentiation of glial cells in vivo are ineffective due to the complex in vivo environment, which hinders the function of factors screened in vitro.

Innovation Solution

An expression vector comprising a nucleic acid sequence encoding an artificial transcription factor with an RZFD core domain, regulated by a glial cell-specific promoter and enhanced by WPRE, capable of binding to RE1, is used to promote the transdifferentiation of non-neuronal cells into functional neurons.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If factors screened in vitro are used to induce neuronal transdifferentiation, then transdifferentiation can be achieved in vitro culture systems, but these factors cannot function effectively in vivo due to the complex in vivo environment

Engineering Contradiction:

Improvetransdifferentiation efficacyVSAvoidenvironmental adaptability

Core Design Contradiction:

ReliabilityVSAdaptability or versatility

Solution Approach 1:

The patent modifies the transcription factor by changing its binding parameters - specifically creating artificial transcription factors with altered DNA binding specificities that recognize RE1 elements. This parameter change allows the factors to function effectively in the complex in vivo environment while maintaining their transdifferentiation capability, resolving the contradiction between in vitro efficacy and in vivo adaptability

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent uses artificial transcription factors as intermediaries that bridge the gap between in vitro screening results and in vivo functionality. These engineered factors serve as mediators that can translate the transdifferentiation potential identified in vitro into effective in vivo action by incorporating elements that allow them to navigate and function in the complex in vivo environment

Inventive Principle:

Principle #24Intermediary (Mediator)

2Ease of manufacture

If complex in vivo environment is considered, then the function of screened factors is hindered, but simplifying the environment is not feasible for therapeutic application

Engineering Contradiction:

Improvetherapeutic applicabilityVSAvoidfactor functionality

Core Design Contradiction:

Ease of manufactureVSReliability

Solution Approach 1:

Instead of trying to simplify the in vivo environment or protect the factors from environmental complexity, the patent inverts the approach by engineering the factors themselves to thrive in the complex environment. The artificial transcription factors are designed with enhanced stability and binding specificity that allow them to function effectively in vivo, making therapeutic application feasible without environmental simplification

Inventive Principle:

Principle #13The other way round (Inversion)

3Measurement precision

If artificial transcription factors with RZFD core domain are used, then specific binding to RE1 is achieved, but ensuring sufficient expression levels in target cells presents a challenge

Engineering Contradiction:

Improvebinding specificityVSAvoidtranscription factor expression level

Core Design Contradiction:

Measurement precisionVSQuantity of substance

Solution Approach 1:

The patent employs preliminary action by using glial cell-specific promoters to drive expression of the artificial transcription factors directly in the target glial cells. This preliminary positioning of the expression system ensures that sufficient quantities of the high-specificity factors are produced in situ, resolving the contradiction between binding precision and expression quantity

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent applies local quality by using cell-type-specific promoters (glial cell-specific promoters) to ensure that the artificial transcription factors are expressed specifically in glial cells where they are needed. This localized expression strategy ensures both high binding specificity to RE1 elements in the correct cell type and sufficient local concentration to drive effective transdifferentiation

Inventive Principle:

Principle #3Local quality

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The expression vector effectively induces the transdifferentiation of glial cells into neurons in vivo, addressing the challenge of in vivo efficacy and providing functional neuronal regeneration.

Implementation Method 1

a nucleic acid sequence encoding an artificial transcription factor capable of binding to RE1, said artificial transcription factor comprises RZFD core domain

Methodology Applied

Scientific EffectDNA binding:

Data Source

PatentEP4692360A1Expression vector and composition for treating nervous system diseases

Publication Date: 2026.02.11 SHANGHAI GENEMAGIC BIOSCIENCES CO LTD

Figure 1A~1B
Figure 1C~1D
Figure 2~3A

AI summary

Provided is an expression vector, comprising: (a) a nucleic acid sequence encoding an artificial transcription factor capable of binding to RE1, the artificial transcription factor comprising an RZFD core domain and (b) a regulatory element for regulating the nucleic acid expression of the artificial transcription factor, wherein the amino acid sequence of the RZFD core domain contains any 5-8 sequences among SEQ ID NOs: 3-10 and does not contain sequences shown in SEQ ID NOs: 12 and 13; the amino acid sequence of the RZFD core domain contains any 6-8 sequences among SEQ ID NOs: 3-10.