S.O.W. ActiveMotif_ChIP-seq_.pdf

Statement of Work

Title: ChIP-seq to identify new transcription factors necessary for embryonic reproductive organ development.

Background: Defects in normal development during gestation can lead to disease states later in life.

Specific cell signaling pathways must be fully explored to understand how and why these types of diseases can occur. One method widely employed by the scientific community, ChIP-seq, is used to discern which genes make up these pathways and how they interact. ChIP-seq can identify areas of chromosomes which are bound by specific proteins and, thus, show different genes working together to produce an affect. By helping to map these pathways and build on the current knowledge of development of reproductive organs, we can increase the chance that diseases will be identified early on and treatments can be found.

The Reproductive Developmental Biology Group has successfully used the ChIP-seq method in the past to identify genes which are critical to the development of testes and ovaries using mouse models. For example, the transcription factor FOXL2 has been linked to some human ovarian developmental diseases and required more study. The Yao Lab has published several manuscripts detailing the importance of

FOXL2 in keeping the ovary on the correct developmental pathway (Nat Commun. 2019 Nov

11;10(1):5116. doi: 10.1038/s41467-019-13060-1., Hum Mol Genet. 2018 Dec 15;27(24):4273-4287. doi:

10.1093/hmg/ddy312., and Sex Dev. 2014;8(5):243-51. doi: 10.1159/000358072. Epub 2014 Jan 29.).

Without a normal functioning FOXL2 protein the ovary changes into a testis. We now would like to continue these studies to further characterize the roles of other critical reproductive genes we have identified.

Scope of Work: The researcher will provide the starting material in the format required by the contractor.

This includes experimental samples and antibodies as available. The contractor shall provide all other resources necessary to accomplish the tasks and deliverables described in this Statement of Work. The contractor shall provide sequencing data and perform quality control analysis at each step of the process.

Required Services: The contractor will prepare chromatin from the samples supplied by the government.

Samples will be frozen primary embryonic gonads, either testes or ovaries. The contractor will do quality control analysis on antibodies as necessary. The contractor will isolate chromatin from the samples, perform

ChIP reactions, construct ChIP-seq libraries, and perform sequencing of those libraries.

Included are all costs for consumable materials such as molecular biology reagents for library construction and sequencing, disposable items (such as pipets, tubes etc.), and cryo-banking of prepared libraries.

Deliverables:

Bioinformatic data specific to four different genes with appropriate quality controls steps during the experiment through to data acquisition.

Period of Performance: September 1, 2020 – August 31, 2021

Place of Performance: Contractor’s facilities.