# Core B

> **NIH NIH P01** · EMORY UNIVERSITY · 2022 · $463,844

## Abstract

The projects within this PPG proposes aims that will determine the transcriptional and epigenetic programs of
plasma cell development and maintenance. Additionally, the projects propose to use genome engineering to
genetically manipulate B cells using CRISPR and over express cDNAs. To provide this expertise, ensure
standardized protocols, integration of results and their subsequent analyses, and the sharing of data, the creation
of an Epigenomics, Bioinformatics, and Genome Engineering Core (Core B) within this PPG is proposed. Core
B will provide state-of-the-art technologies, molecular biology expertise, and bioinformatic services that assess
DNA methylation, chromatin state and accessibility, and transcript expression through deep sequencing of both
bulk and single-cell datasets. To support the genome engineering experiments Core B will identify functional
sgRNA, provide cloning and viral preparation services, maintain plasmid repository and protocols supporting B
cell genome engineering. To serve the projects, three Aims are proposed. Aim 1. Provide uniform and quality
library preparation and sequencing to determine the transcriptome, DNA methylation patterns,
chromatin accessibility, and histone modifications. Core B will create high-quality libraries and facilitate
deep sequencing based on five technologies to derive epigenetic programming. RNA-seq will be used to
determine the transcriptome. Reduced Representation Bisulfite Sequencing (RRBS) or Whole Genome Bisulfite
Sequencing (WGBS) will be used to assess DNA methylation. The Assay for Transposase Accessible Chromatin
(ATAC-seq) will determine chromatin accessibility. Cleavage Under Targets and Tagmentation (CUT&Tag) will
be used to determine histone posttranslational modifications. Aim 2. Provide iterative bioinformatic
computational analysis of datasets. A question driven, iterative bioinformatics analysis will be used to derive
and examine the molecular programming of B cells and plasma cells. Core B will draw upon considerable
expertise in both single-cell and bulk B cell/plasma cell genomic datasets with the capability of integrating data
across platforms, disease and conditions, and species. Core B will provide long-term data storage, and facilitate
sharing of processed datasets, including the use of interactive data exploration tools to facilitate data analysis
by the entire program. Aim 3. Provide a B cell genome engineering platform using CRISPR/Cas9 and cDNA
overexpression. Core B will test sgRNAs to identify those that provide maximal deletion, clone sgRNAs of
interest into viral-based vectors and prepare stocks, and provide optimized protocols for infection and ultimately
genome engineering of B cells. Additionally, Core B will maintain a centralized repository of vectors that contain
flow cytometry compatible markers for sorting and selection, genome-wide sgRNA pools, and constructs that
allow overexpression of cDNAs. Thus, Core B will provide a common library preparation an...

## Key facts

- **NIH application ID:** 10428166
- **Project number:** 2P01AI125180-06
- **Recipient organization:** EMORY UNIVERSITY
- **Principal Investigator:** Christopher D Scharer
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $463,844
- **Award type:** 2
- **Project period:** 2016-06-25 → 2027-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10428166

## Citation

> US National Institutes of Health, RePORTER application 10428166, Core B (2P01AI125180-06). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10428166. Licensed CC0.

---

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