# Core 1: Expression and Molecular Biology (EMB)

> **NIH NIH P01** · UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB · 2023 · $725,754

## Abstract

SUMMARY- CORE 1: EXPRESSION AND MOLECULAR BIOLOGY (EMB) CORE
High quality protein samples are rate limiting for most structural and mechanistic investigations of DNA Repair
(DR) complexes and assemblies. Core 1, the Expression and Molecular Biology (EMB) Core, is designed to
lower these barriers for individual labs by providing access to centralized resources and expertise in advanced
recombinant protein technologies. We will implement modern DNA assembly technologies and incorporate
efficient protein production strategies to overcome these technical bottlenecks. The EMB Core will serve as a
research, production, and training resource for all four Projects. We will develop robust pipelines for (1)
engineering vectors to aid soluble expression and purification for three established expression systems –
bacteria, insect cells and human cells, and constructing single-gene and multi-gene expression vectors using
advanced DNA assembly techniques, such as BioBrick and Gibson assembly, (2) providing expression systems
that facilitate efficient purification of well-behaved, functional DR complexes, e.g. by incorporating cleavable GFP
tags to be employed with GFP-binder single-chain nanobody columns, (3) validating protein interactions and
partnerships in DR complexes and assemblies, and (4) using streamlined scaled-up expression platforms to
support large-scale cell production. We propose three Aims to empower SBDR-5.
Aim 1. Meet the molecular biological needs of SBDR. We will generate in-house modified vectors and create
expression constructs for expressing single DR proteins and multi-protein complexes for desired host systems.
Aim 2. Provide key protein production resources for SBDR. The available resources include large-scale insect
cell and bacteria production, selected human cell production for difficult-to-express DR targets, purification of
recombinant proteins when needed, and validation of interacting partners in protein complexes.
Aim 3. Establish and maintain a Repository/Distribution Center for SBDR. We have amassed a large collection
of key reagents and maintained a Repository/Distribution Center. A secure EMB website with a searchable
database will be created to streamline access to these resources and allow timely distribution of the requested
reagents to the SBDR labs.
All four Projects will rely on EMB Core services to support their molecular biological and protein needs. The EMB
Core has established close interactions with all Projects during the previous four funding cycles. Ready access
to EMB Core’s enabling expertise and resources will synergize all four Projects, avoid duplication of research
efforts, and promote active collaborations. The proposed EMB technologies and services will, in concert with the
SCB Core, accelerate SBDR-5 research and empower SBDR Projects by facilitating quantitative, high-resolution
structural insights for DNA repair machinery functions.

## Key facts

- **NIH application ID:** 10695057
- **Project number:** 5P01CA092584-23
- **Recipient organization:** UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
- **Principal Investigator:** Miaw-Sheue Tsai
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $725,754
- **Award type:** 5
- **Project period:** 2001-09-27 → 2026-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10695057

## Citation

> US National Institutes of Health, RePORTER application 10695057, Core 1: Expression and Molecular Biology (EMB) (5P01CA092584-23). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10695057. Licensed CC0.

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