Cell-Specific Paired Interrogation of the Mouse Ovarian Epigenome and Transcriptome.
J Vis Exp
; (192)2023 02 24.
Article
em En
| MEDLINE
| ID: mdl-36912526
ABSTRACT
Assessing cell-type-specific epigenomic and transcriptomic changes are key to understanding ovarian aging. To this end, the optimization of the translating ribosome affinity purification (TRAP) method and the isolation of nuclei tagged in specific cell types (INTACT) method was performed for the subsequent paired interrogation of the cell-specific ovarian transcriptome and epigenome using a novel transgenic NuTRAP mouse model. The expression of the NuTRAP allele is under the control of a floxed STOP cassette and can be targeted to specific ovarian cell types using promoter-specific Cre lines. Since recent studies have implicated ovarian stromal cells in driving premature aging phenotypes, the NuTRAP expression system was targeted to stromal cells using a Cyp17a1-Cre driver. The induction of the NuTRAP construct was specific to ovarian stromal fibroblasts, and sufficient DNA and RNA for sequencing studies were obtained from a single ovary. The NuTRAP model and methods presented here can be used to study any ovarian cell type with an available Cre line.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Transcriptoma
/
Epigenoma
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article