# Development of Compact CRISPR Editors for Multiplexed Editing of Complex Gene Networks

> **NIH NIH DP2** · BOSTON COLLEGE · 2020 · $151,805

## Abstract

Development of Compact CRISPR Editors for Multiplexed Editing of Complex
Gene Networks
Abstract
 O-Linked N-acetylglucosamine (O-GlcNAc) is a monosaccharide modification of
nucleocytoplasmic proteins that serves as an important chemical code in regulation and
signaling processes. Specifically, O-GlcNAc modification of transcription factors plays a
profound role in transcription regulation. However, previous efforts in studying the dynamic
O-GlcNAc code in a complex gene network were hampered by the inability to perform
protein-specific and locus-specific O-GlcNAc modification of transcription factors. The
objective of our parental 1DP2HG011027-01 project is to develop compact, multiplexable
CRISPR editors to allow the genomic editing at transcriptional, epigenetic, and post-
translational levels. In line with this parental project, we now request an administrative
supplement with the objective to extend our CRISPR editors to enable locus-specific
engineering of the O-GlcNAc modification of transcription factors and study how dynamic
O-GlcNAc codes regulate transcription and gene expression. To achieve this objective,
we will collaborate with Professor Christina Woo’s group at Harvard University and adopt
the tools developed by their 1U01CA242098-01 project in our efforts to develop CRISPR
editors for engineering transcription factor-specific O-GlcNAc modifications. By fusing O-
GlcNAc transferase (OGT) and O-GlcNAcase (OGA) with a nanobody, Woo’s O-GlcNAc
engineering constructs can be efficiently recruited by Cas9 proteins tagged by the epitope
of the nanobody. We will investigate the optimal distance between the CIRPSR-mediated
OGT/OGA binding site and the target transcription factor using specifically designed
reporter constructs that translate the transcription factor activity into reporter signal in vivo.
Furthermore, we will integrate the RNA aptamer technology developed in our parental
project into proximity-directed O-GlcNAc engineering and evolve RNA aptamers in vitro
and in vivo that can recruit endogenous OGT and OGA to the CRISPR editors for
reprogramming O-GlcNAc codes on target transcription factors. The proposed research
will lead to a novel method to engineer the O-GlcNAc code of transcription factors and
allow us to study the dynamic interplay of O-GlcNAc modifications at different transcription
factors in gene networks.

## Key facts

- **NIH application ID:** 10136340
- **Project number:** 3DP2HG011027-01S1
- **Recipient organization:** BOSTON COLLEGE
- **Principal Investigator:** Jia Niu
- **Activity code:** DP2 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $151,805
- **Award type:** 3
- **Project period:** 2020-08-07 → 2022-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10136340

## Citation

> US National Institutes of Health, RePORTER application 10136340, Development of Compact CRISPR Editors for Multiplexed Editing of Complex Gene Networks (3DP2HG011027-01S1). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10136340. Licensed CC0.

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