Investigating the role of an RNA-binding protein family in mammalian infertility
Lead Research Organisation:
University of Edinburgh
Department Name: MRC Centre for Reproductive Health
Abstract
Although infertility affects 10-15% of couples worldwide, its genetics remains poorly understood. However, it is clear that germ cell development and function relies on the exquisite regulation of gene expression, including at the level of mRNA translation and stability. Cytoplasmic RNA-binding proteins, of which there are > 1,500 in man are key regulators of mRNA translation and stability and consequently their loss of function can result in infertility.
Poly(A)-binding proteins (PABPs) are an RNA-binding protein family, which bind to the 3'poly(A) tail of mRNAs, and act as central regulators of mRNA fate. Whilst PABPs are known to be critical for fertility in invertebrates, their roles in mammalian fertility are less clear. However, several lines of evidence support a key role. Firstly, changes in poly(A) tail length are a key mechanism for regulating gene expression in germ cells from worms to man. Secondly, PABPs are critical for the function of other RNA-binding protein families required for fertility in mouse and man. Thirdly, changes in available PABP levels disrupts gametogenesis in male mice. Consistent with this indirect evidence knock-out of embryonic PABP in mice results in female infertility.
However, it is clear that other PABP family members must also be playing a vital role (e.g. based on expression patterns, timing of poly(A) tail length changes, and phenotypic comparisons), but an absence of knock-out mouse models has prevented their investigation. This project addresses this important knowledge gap utilising newly generated knock-out mice. The student will characterise the fertility of these mice (male and female) and establish the changes that underlie observed changes in fertility. As we have multiple PABP knock-out conditional mice, mice with cell type specific loss of PABPs or compound mice (i.e. missing more than one PABP) can be generated to further investigate their roles in fertility. As part of their analysis, students may also uncover additional exciting reproductive or non-reproductive phenotypes.
This project will utilise a variety of techniques including mouse genetics, mouse phenotype analysis, histology, immunohistochemistry, RT-PCR, qRT-PCR, western blotting, ELISA and hormone assays. Other techniques will be employed as lead by results.
The laboratory has several post-docs, PhD students and research support staff and utilises a highly-interdisciplinary approach (e.g. combining work in model organisms, in vitro manipulation of germ cells, molecular biology, biochemistry and biophysics).
Poly(A)-binding proteins (PABPs) are an RNA-binding protein family, which bind to the 3'poly(A) tail of mRNAs, and act as central regulators of mRNA fate. Whilst PABPs are known to be critical for fertility in invertebrates, their roles in mammalian fertility are less clear. However, several lines of evidence support a key role. Firstly, changes in poly(A) tail length are a key mechanism for regulating gene expression in germ cells from worms to man. Secondly, PABPs are critical for the function of other RNA-binding protein families required for fertility in mouse and man. Thirdly, changes in available PABP levels disrupts gametogenesis in male mice. Consistent with this indirect evidence knock-out of embryonic PABP in mice results in female infertility.
However, it is clear that other PABP family members must also be playing a vital role (e.g. based on expression patterns, timing of poly(A) tail length changes, and phenotypic comparisons), but an absence of knock-out mouse models has prevented their investigation. This project addresses this important knowledge gap utilising newly generated knock-out mice. The student will characterise the fertility of these mice (male and female) and establish the changes that underlie observed changes in fertility. As we have multiple PABP knock-out conditional mice, mice with cell type specific loss of PABPs or compound mice (i.e. missing more than one PABP) can be generated to further investigate their roles in fertility. As part of their analysis, students may also uncover additional exciting reproductive or non-reproductive phenotypes.
This project will utilise a variety of techniques including mouse genetics, mouse phenotype analysis, histology, immunohistochemistry, RT-PCR, qRT-PCR, western blotting, ELISA and hormone assays. Other techniques will be employed as lead by results.
The laboratory has several post-docs, PhD students and research support staff and utilises a highly-interdisciplinary approach (e.g. combining work in model organisms, in vitro manipulation of germ cells, molecular biology, biochemistry and biophysics).
Organisations
People |
ORCID iD |
| Nicola Gray (Primary Supervisor) | |
| Hristina Gyurova (Student) |