# A novel epigenetic mechanism in early embryogenesis

> **NIH NIH R35** · YALE UNIVERSITY · 2023 · $193,855

## Abstract

Our parental proposal aims to investigate critical mechanisms in mammalian early
embryogenesis, such as endogenous transposable elements (TEs) remnants of ancient
transposons and DNA methylation. We use embryonic stem cell (ESC) culture, genetic-
engineered mouse model and in vitro fertilization (IVF) mouse models to interrogate the
transcription and epigenetic regulatory pathways in ESC or developing mouse fetus.
 The current proposal advocates the needs for purchasing a versatile real-time PCR
machine to accurately quantify the gene expression, epigenetic factors and genotype in early
embryonic and extraembryonic tissues (Aim 1 and 2) and run protein thermal shift assays for
biochemistry experiments (Aim3).
 Nearly half of our samples are generated from limited amounts of materials, such as
100-200 cells from morula or blastocyst stage mouse fetus. Our targets are often challenging,
such as TEs that are highly repetitive. These unique research needs demand a precise and
reliable instrument which also has the capability to run the Taqman assays as an orthogonal
approach to independently validate the results from SYBR green-based real-time PCR assays.
Furthermore, our experiments often require different throughputs, which need to interchange
between a 96-well and 384-well plates. For example, a routine examination of gene expression
level in embryonic stem cells needs a routine SYBR-green based assay in 96-well plates,
whereas an examination of TE expression levels in mouse blastocyst needs Taqman assays in
384-well plates.
 Moreover, a major direction of our parental proposal is to investigate the function of
SATB1, a critical remodeling factors binding to DNA, as well as novel DNA methyltransferases
(Aim3). Thus, we need to an instrument that can run protein thermal shift assays to check
protein quality and their binding to DNA substrates as shown in our recently published studies.
 This proposal aims to acquire an accurate, versatile and fast Real-Time PCR machine
that meets multiple unique research needs in the parental proposal outlined above. The
outcome of the proposal will greatly facilitate our investigation in epigenetics and
embryogenesis.

## Key facts

- **NIH application ID:** 10799233
- **Project number:** 3R35GM136346-04S1
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** zhuo Andrew Xiao
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $193,855
- **Award type:** 3
- **Project period:** 2020-04-01 → 2025-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10799233

## Citation

> US National Institutes of Health, RePORTER application 10799233, A novel epigenetic mechanism in early embryogenesis (3R35GM136346-04S1). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10799233. Licensed CC0.

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