Overview

The Functional Genomics Shared Resource provides reagents, infrastructure, and expertise for high-throughput screening of chemical, CRISPR, RNAi, and open reading frame (ORF) libraries.

The screening platforms supported by the facility enable systematic, comprehensive interrogation of gene function or molecular processes that provide opportunities for the discovery of new targets, lead compounds, or biological insights. In addition, the shared resource provides custom cell line engineering services using CRISPR techniques.

Services

The shared resource provides services in three major areas: compound screening, functional genetic screening, and custom genetic perturbation. Instrumentation available for quantitative assays includes multimodal plate readers and the ThermoFisher CellInsight CX5 High-Content Screening (HCS) system.

Compound Screening

The shared resource assists investigators perform high-throughput compound screens in 96- or 384-well format using screener-provided or in-house libraries. In-house libraries range in size from 120 to over 50,000 compounds. More information.

Functional Genomic Screening

Libraries available for loss-of-function screening include human siRNA (arrayed), lentiviral human shRNA (arrayed or pooled), lentiviral human and mouse CRISPR/CRISPRi sgRNA (pooled). Libraries for gain-of-function screening include lentiviral human kinome ORFs (arrayed or pooled), lentiviral human ORFs (pooled) or CRISPRa lentiviral human CRISPR sgRNAs (pooled). Services to generate custom CRISPR libraries for pooled and arrayed screening are also available.

Custom genetic perturbation solutions for functional analysis of individual genes

This shared resource can work with investigators to create knockdown, knockout, knock-in or overexpression cell lines using shRNA-, CRISPR- or ORF-based approaches. Individual lentiviral shRNAs from the Sigma Mission human collection are also available for distribution.

Equipment

Labcyte Echo Acoustic Dispenser for fast and flexible transfers of nanoliter volumes, ideal for compound screening and dose-response experiments. This instrument is available for scheduling through CoreResearch@Duke.

Bravo Automated Liquid Handler for high-throughput transfers of microliter volumes for 96- or 384-well plates integrated with barcode labeler, heat sealer and BenchCel Robot for microplate management. The entire system is housed in a Baker BIOPROtect II cabinet.

CellInsight CX5 High Content Screening (HCS) system for automated acquisition and analysis of plate-based images, integrated with robotic plate stackers and barcode reader for high-throughput plate processing. This instrument is available for scheduling through CoreResearch@Duke.

BMG Clariostar multimodal plate reader with detectors for fluorescence intensity, luminescence, absorbance, FP, TRF, FRET and TR-FRET. This reader is also equipped with dual injectors, temperature control to 45oC, barcode readers and stackers for high-throughput plate processing. This instrument is available for scheduling through CoreResearch@Duke.

Biotek ELx405 microplate washer, Biotek uFill dispenser, Matrix Wellmate microplate dispenser and VP aspirating wands and manifolds for fast and accurate high-throughput plate processing. Biotek MultiFlo microplate dispenser for automated filling of plates.

Tissue Culture facility, fully equipped with two 6’ biosafety cabinets, six CO2 incubators, EVOS fluorescence microscope, tabletop centrifuges, water bath, freezers and refrigerators, dedicated for use by screeners.

Services and Fees

Services and resources provided by the shared resource are available to all DCI members. The Functional Genomics Shared Resource does not currently maintain a fee schedule for DCI members but offers priority access to these services to DCI members. Members interested in utilizing this resource should contact So Young Kim, PhD.

Location

BOX 3053
0050 CARL Building
213 Research Drive
Durham, NC