# Regulation of X-chromosome inactivation and imprinting by non-coding elements

> **NIH NIH R01** · MASSACHUSETTS GENERAL HOSPITAL · 2024 · $192,248

## Abstract

PROJECT SUMMARY
Interactions between long noncoding RNAs and epigenetic complexes are now increasingly recognized as
important regulators of developmental decisions. A classic example is X-chromosome inactivation (XCI), an
epigenetic process by which one X-chromosome becomes inactivated in the early embryo. XCI can take
place randomly or in an imprinted manner on the paternal X. In the mouse, random XCI is observed in the
embryo proper (soma), and imprinted XCI in the placenta. Both require the ‘X-inactivation center’ (Xic), an
X-linked region that harbors several essential noncoding genes. A key gene is Xist, which produces a
noncoding RNA that ‘paints’ the X-chromosome and recruits silencing factors in cis. Our work has shown
that, during random XCI, Xist is regulated both positively and negatively by Xic factors, with Jpx in the
positive pathway and Tsix/Xite in the negative pathway. We have also shown that homologous X-
chromosome pairing coordinates which X chromosome will be inactivated. In this funding cycle, we have
further advanced understanding of the Xic’s role in X-chromosome counting, allelic choice, and imprinting.
For the counting mechanism, Jpx RNA — an Xist activator — serves as an X-linked “numerator” by titrating
the autosomal “denominator”, CTCF. This titration accounts for the X-to-autosome (X:A) ratio and creates a
permissive state for Xist induction. We also learned that X-X pairing requires a new long noncoding RNA
called “PAR-TERRA” produced from the X-linked pseudoautosomal region (PAR). Interestingly, all pairing
interactions take place within a “tetrad” of two Xic’s and two PAR’s. X-X pairing is then resolved by mRNA
decapping enzyme, DCP1A. For imprinted XCI, we have generated an ex vivo cellular model to study XP
imprinting. We have demonstrated that a 200-kb Xic region is sufficient to direct Xist imprinting and that Xist
imprinting requires an unpaired state in the male germ line that is akin to meiotic sex chromosome
inactivation (MSCI). Our research has generated new opportunities and many additional mechanistic
questions. In the next funding cycle, we aim to (i) investigate mechanisms underlying X-X pairing and
breaking of X-chromosome symmetry, (ii) determine how Jpx-CTCF titration regulates the initiation of XCI,
and (iii) elucidate the mechanisms of imprinted XCI. The proposed work will require five years to complete
and is expected to train three postdoctoral fellows for academic careers in the fields of epigenetics and RNA
biology.

## Key facts

- **NIH application ID:** 11094463
- **Project number:** 3R01GM058839-26S1
- **Recipient organization:** MASSACHUSETTS GENERAL HOSPITAL
- **Principal Investigator:** JEANNIE T LEE
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $192,248
- **Award type:** 3
- **Project period:** 1999-01-01 → 2026-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/11094463

## Citation

> US National Institutes of Health, RePORTER application 11094463, Regulation of X-chromosome inactivation and imprinting by non-coding elements (3R01GM058839-26S1). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/11094463. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*