1. Barhl 1 is required for the differentiation of inner ear hair cell-like cells from mouse embryonic stem cells
- Author
-
Jinfu Wang, Zhenhuang Chen, Hui Jiang, Jian-Zhong Shao, Liquan Huang, Min-Xin Guan, Xiao Huang, Cuicui Wang, Chao Zhong, and Xiao-Cui Luo
- Subjects
0301 basic medicine ,Nerve Tissue Proteins ,Deafness ,Biology ,medicine.disease_cause ,Biochemistry ,Cell Line ,Mice ,03 medical and health sciences ,0302 clinical medicine ,otorhinolaryngologic diseases ,medicine ,Animals ,Inner ear ,Progenitor cell ,Gene ,Mutation ,Hair Cells, Auditory, Inner ,Hair cell differentiation ,integumentary system ,Cell Differentiation ,Mouse Embryonic Stem Cells ,Cell Biology ,Embryonic stem cell ,Cell biology ,Repressor Proteins ,030104 developmental biology ,medicine.anatomical_structure ,Homeobox ,sense organs ,Hair cell ,CRISPR-Cas Systems ,Gene Deletion ,030217 neurology & neurosurgery - Abstract
Inner ear hair cells are mechanoreceptors responsible for hearing. Pathogenic defects of hair cell-specific genes are one of the major causes of deafness. The BarH class homeobox gene Barhl1 is a deafness gene expressed in developing hair cells, yet the role of Barhl1 during hair cell development remains poorly understood. In the present study, we first established an in vitro differentiation system to efficiently obtain mouse embryonic stem cell (mESC)-derived hair cell-like cells. Subsequently, a mESC line carrying a targeted disruption of Barhl1 was generated using CRISPR/Cas9 technology and subjected to the established in vitro hair cell differentiation protocol. Targeted disruption of Barhl1 does not affect the induction of mESCs toward early primitive ectoderm-like (EPL) cells and otic progenitors but strongly inhibits the differentiation of hair cell-like cells. Using RNA-sequencing and bioinformatics, we further unravel the molecular mechanism underlying Barhl1-mediated hair cell development. Our data demonstrate the essential role of Barhl1 during hair cell development and provide a basis for the treatment of Barhl1 mutation-based deafness.
- Published
- 2018