Humanized skeletal muscle

Patent metadata

Field	Value
Publication number	US-10874092-B2
Application number	US-201615739042-A
Country	US
Kind code	B2
Filing date	Jun 30, 2016
Priority date	Jun 30, 2015
Publication date	Dec 29, 2020
Grant date	Dec 29, 2020

Abstract

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.

First claim

What is claimed is:

1. A chimeric pig embryo comprising: (i) homozygous disruptions of endogenous MYF5, MYOD, and MRF4 genes in it genome, wherein said disruptions result in no expression of the endogenous MYF5, MOYD, and MRF4 proteins; and (ii) human skeletal muscle cells expressing human MYF5, MYOD, and MRF4 proteins, wherein the human skeletal muscle cells are differentiated from human pluripotent stem cells introduced into a pig embryo at an earlier stage of development than said chimeric pig embryo and wherein the introduced human pluripotent cells integrated into the inner cell mass of the pig embryo at the blastocyst stage.

2. A chimeric pig blastocyst comprising: (i) homozygous disruptions of endogenous MYF5, MYOD, and MYF4 genes in its genome, wherein said disruptions result in no expression of the endogenous MYF5, MOYD, and MRF4 proteins; and (ii) human pluripotent cells having intact MYF5, MYOD, and MRF4 genes, wherein the human pluripotent cells integrate into the inner cell mass of the blastocyst.

Assignees

• Univ Minnesota

Inventors

• Garry Daniel J
• Garry Mary G
• Koyano Naoko

Classifications

• A01K2217/075

  inducing loss of function, i.e. knock out · CPC title

• A01K2267/025

  Animal producing cells or organs for transplantation · CPC title

• A61K35/34

  Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes (vascular smooth muscle A61K35/44) · CPC title

• A01K2217/00

  Genetically modified animals · CPC title

• C12N5/0604

  Whole embryos; Culture medium therefor · CPC title