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DOI: 10.3791/64765-v
Sarah R. Ocañas*1,2,3, José V. V. Isola*1,3, Tatiana D. Saccon1, Kevin D. Pham2, Ana J. Chucair-Elliott2, Augusto Schneider4, Willard M. Freeman2,3, Michael B. Stout1,3
1Aging and Metabolism Research Program,Oklahoma Medical Research Foundation, 2Genes & Human Disease Research Program,Oklahoma Medical Research Foundation, 3Oklahoma City Veterans Affairs Medical Center, 4Nutrition College,Federal University of Pelotas
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This study optimizes the translating ribosome affinity purification (TRAP) and isolation of nuclei tagged in specific cell types (INTACT) methods for paired interrogation of the ovarian transcriptome and epigenome in a mouse model. The approach enables analysis of specific ovarian cell populations without cell sorting, facilitating insights into cell type-specific changes associated with aging.
在该协议中,翻译核糖体亲和纯化(TRAP)方法和特定细胞类型(INTACT)方法中标记的细胞核的分离针对使用NuTRAP小鼠模型交叉到Cyp17a1-Cre小鼠系的细胞特异性卵巢转录组和表观基因组的配对询问进行了优化。
该方法提供了一种从同一只小鼠的特定卵巢细胞群中分离核酸的技术,而无需细胞分选。该技术可以帮助确定特定细胞群在各种条件下的变化。例如,随着年龄的增长。
该技术的主要优点是它允许对特定卵巢细胞类型进行配对的表观基因组和转录组学分析,而无需细胞分选。这显著提高了通量,降低了成本,并且在对卵巢组织进行细胞类型特异性分析时无需专用设备。在这里,我们的目标是确定生殖衰老前卵巢衰老发生的细胞类型特异性变化,这对治疗女性不孕症具有意义。
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