Humanized Skeletal Muscle via MYF5 Knockout Pigs
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Solution Overview
Problem
Current treatments for terminal muscle diseases are inadequate, and there is a lack of therapies for progressive muscle weakness associated with aging, with limited options for transplantation due to donor organ availability and immune rejection concerns.
Innovation Solution
Development of MYF5/MYOD/MRF4 knockout pigs or other animals as hosts for producing personalized humanized muscle cells through gene editing technologies, creating a niche for human stem cells to generate humanized skeletal muscle, thereby addressing immune rejection and organ availability issues.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If human stem cells are transplanted directly into patients, then muscle tissue can be replaced, but immune rejection occurs
Solution Approach 1:
The patent uses a genetically modified non-human animal (with deleted MYF5, MYOD, and MRF4 genes) as an intermediary host. This host lacks the ability to form functional skeletal muscle, creating a permissive niche that accepts human stem cells without triggering strong immune rejection, while still providing the necessary physiological environment for muscle regeneration.
Solution Approach 2:
The patent fundamentally changes the genetic parameters of the host animal by deleting three critical myogenic regulatory genes (MYF5, MYOD, MRF4). This genetic modification transforms the host from an immunologically incompatible species into a permissive environment that supports human stem cell engraftment and differentiation into functional human skeletal muscle tissue.
2Quantity of substance
If donor organs are used for transplantation, then muscle tissue replacement is possible, but donor organ availability is limited
Solution Approach 1:
The genetically modified animal host serves itself by providing the necessary physiological environment and resources for human stem cell differentiation into functional muscle tissue. The host's body systems (circulation, nutrition, innervation) automatically support the development of humanized muscle without requiring external donor organs or intensive human intervention.
Solution Approach 2:
The patent performs preliminary genetic modification of the animal host before stem cell transplantation. By pre-deleting the MYF5, MYOD, and MRF4 genes, the host is prepared in advance to accept and support human stem cells, eliminating the need to search for compatible human donors and significantly increasing tissue availability.
3Adaptability or versatility
If gene editing is performed on multiple myogenic genes, then a permissive niche is created, but the complexity of the procedure increases
Solution Approach 1:
The patent divides the complex task of creating a permissive niche into three separate gene targeting events (MYF5, MYOD, MRF4). Each gene can be edited independently using standard CRISPR/Cas9 or TALEN techniques, allowing the complex modification to be achieved through multiple simple, modular steps rather than one complex procedure.
Solution Approach 2:
The patent uses universal gene editing tools (CRISPR/Cas9 or TALEN systems) that can target multiple different genes with the same basic methodology. This multi-functional approach allows the same technical platform to delete MYF5, MYOD, and MRF4 genes, reducing the need for different specialized techniques and simplifying the overall procedure.
Data Source
AI summary
Described herein is a method for producing a chimeric non-human animal expressing a human a MYF5, MYOD, MRF4 gene or a combination thereof gene comprising: a) generating an MYF5, MYOD, MRF4 or combination thereof null non-human animal cell, wherein both copies of the non-human MYF5, MYOD, MRF4 gene or combination thereof carry a mutation that prevents production of functional MYF5, MYOD, MRF4 protein or combination thereof in said non-human animal; b) creating a MYF5, MYOD, MRF4 or combination thereof null non-human blastocyst by somatic cell nuclear transfer comprising fusing a nucleus from said MYF5, MYOD, MRF4 or combination thereof null non-human animal cell of a) into an enucleated non-human oocyte and activating said oocyte to divide so as to form an MYF5, MYOD, MRF4 or combination thereof null non-human blastocyst; c) introducing human stem cells into the MYF5, MYOD, MRF4 or combination null non-human blastocyst of b); and d) implanting said blastocyst from c) into a pseudopregnant surrogate non-human animal to generate a chimeric non-human animal expressing human MYF5, MYOD, MRF4 or combination thereof.