# Core 1: Expression and Molecular Biology (EMB) Core

> **NIH NIH P01** · UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB · 2020 · $436,668

## Abstract

Core 1 – Expression and Molecular Biology (EMB) Core
PROJECT SUMMARY/ABSTRACT
The overall goal of SBDR-4 is to characterize the DNA repair multi-protein machines and assemblies that
maintain genomic integrity and to identify the mechanisms that underlie detection and signaling of the damage,
in order to recruit repair machines to the damage site for repair. The proposed SBDR research will provide
mechanistic, predictive biology for cancer interventions through a framework built upon structural and
functional understanding of DNA repair machines. The major challenges faced by SBDR-4 include: (1) efficient
reconstitution and assembly of full-length and modified proteins and complexes that control integrity of the
genome, (2) determining the solution structures of flexible multi-protein complexes that are spatially and
temporally regulated in cells, and (3) linking structures to biochemistry and cellular phenotypes. The
Expression and Molecular Biology (EMB) Core will overcome the technical aspects of these bottlenecks by
implementing robust DNA assembly technologies, incorporating efficient protein purification strategies, and
developing advanced tools for cellular studies. The EMB Core services will empower SBDR-4 by providing
high-quality key starting materials for cell biological, biochemical, biophysical, and structural investigations of
DNA repair machines. The EMB Core will serve as a research, production, and development resource for all
five Projects by providing dedicated Core staff and centralized expertise. The EMB Core will develop robust
pipelines for (1) construction of single-gene and multi-gene expression vectors, (2) creation of isogenic human
cell lines with designed changes, (3) streamlined scaled-up expression technologies, (4) purification of well-
behaved, functional DNA repair proteins and complexes, and (5) validation of protein interactions and
partnerships. The innovative technologies provided by EMB Core to enable SBDR-4 research include: (1) high-
efficiency cloning of multi-gene vectors using advanced DNA assembly technologies, e.g. the BioBrick-based
MacroBac system, Gibson assembly and Golden Gate assembly; (2) incorporation of cleavable GFP tags and
application of GFP-binder single-chain nanobody columns for efficient purification of multimeric protein
complexes for in vitro analysis; and (3) developing novel cellular tools using a novel, highly effective CRISPR
knock-in (KI) method to create isogenic human cell lines with desired changes, as well as using the HIV-TAT
methodology to produce interfering peptides for cellular studies. The EMB Core thus offers a robust blend of
established and new technologies and efficiently provides reagents to jump-start Project Aims and cross-
Project interactions for characterizing transient interactions and dynamic conformations that control the
assembly and function of multi-protein complexes in response to DNA damage to maintain genomic stability.

## Key facts

- **NIH application ID:** 10003189
- **Project number:** 5P01CA092584-20
- **Recipient organization:** UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
- **Principal Investigator:** Priscilla K. Cooper
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $436,668
- **Award type:** 5
- **Project period:** 2001-09-27 → 2021-09-20

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10003189

## Citation

> US National Institutes of Health, RePORTER application 10003189, Core 1: Expression and Molecular Biology (EMB) Core (5P01CA092584-20). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10003189. Licensed CC0.

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