PUBLICATION
Heading towards a dead end: The role of DND1 in germ line differentiation of human iPSCs
- Authors
- Mall, E.M., Lecanda, A., Drexler, H.C.A., Raz, E., Schöler, H.R., Schlatt, S.
- ID
- ZDB-PUB-211019-7
- Date
- 2021
- Source
- PLoS One 16: e0258427 (Journal)
- Registered Authors
- Raz, Erez
- Keywords
- none
- MeSH Terms
-
- CRISPR-Cas Systems/genetics
- Cell Differentiation
- Cell Lineage
- Cell Proliferation
- Gene Editing
- Gene Expression
- Germ Cells/cytology
- Germ Cells/metabolism*
- Humans
- Induced Pluripotent Stem Cells/cytology
- Induced Pluripotent Stem Cells/metabolism
- Neoplasm Proteins/genetics
- Neoplasm Proteins/metabolism*
- Positive Regulatory Domain I-Binding Factor 1/genetics
- Positive Regulatory Domain I-Binding Factor 1/metabolism
- Principal Component Analysis
- RNA/chemistry
- RNA/genetics
- RNA/metabolism
- RNA, Guide, Kinetoplastida/metabolism
- RNA-Binding Proteins/genetics
- RNA-Binding Proteins/metabolism
- Single-Cell Analysis
- PubMed
- 34653201 Full text @ PLoS One
Citation
Mall, E.M., Lecanda, A., Drexler, H.C.A., Raz, E., Schöler, H.R., Schlatt, S. (2021) Heading towards a dead end: The role of DND1 in germ line differentiation of human iPSCs. PLoS One. 16:e0258427.
Abstract
The DND microRNA-mediated repression inhibitor 1 (DND1) is a conserved RNA binding protein (RBP) that plays important roles in survival and fate maintenance of primordial germ cells (PGCs) and in the development of the male germline in zebrafish and mice. Dead end was shown to be expressed in human pluripotent stem cells (PSCs), PGCs and spermatogonia, but little is known about its specific role concerning pluripotency and human germline development. Here we use CRISPR/Cas mediated knockout and PGC-like cell (PGCLC) differentiation in human iPSCs to determine if DND1 (1) plays a role in maintaining pluripotency and (2) in specification of PGCLCs. We generated several clonal lines carrying biallelic loss of function mutations and analysed their differentiation potential towards PGCLCs and their gene expression on RNA and protein levels via RNA sequencing and mass spectrometry. The generated knockout iPSCs showed no differences in pluripotency gene expression, proliferation, or trilineage differentiation potential, but yielded reduced numbers of PGCLCs as compared with their parental iPSCs. RNAseq analysis of mutated PGCLCs revealed that the overall gene expression remains like non-mutated PGCLCs. However, reduced expression of genes associated with PGC differentiation and maintenance (e.g., NANOS3, PRDM1) was observed. Together, we show that DND1 iPSCs maintain their pluripotency but exhibit a reduced differentiation to PGCLCs. This versatile model will allow further analysis of the specific mechanisms by which DND1 influences PGC differentiation and maintenance.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping