The UFHCC CRISPR Functional Screening (CSF) developing shared resource provides cancer researchers with access to the expertise and tools to apply CRISPR-based genome editing approaches to enable functional screening of diverse biological systems relevant to cancer research.
The aims of the CFS-DSR are to: 1) provide consultation on study design of CRISPR based functional screening approaches in cancer research; 2) produce screen ready stocks of selected genome wide and targeted screen ready CRISPR libraries in standard vector systems; 3) construct custom CRISPR libraries for investigator defined targeted gene sets; 4) enable functional screening through development of standard protocols(SOPs), training, and technical assistance in implementation of screening methods; 5) work collaboratively with investigators in the development and troubleshooting of novel approaches and innovative methodology at the forefront of screening technologies; 6) coordinate routine NGS and bioinformatic analysis of functional screening data sets.
The CFS supports functional screening by cancer researchers through expert consultation, resource development and project/team specific assistance. CFS provides both the capabilities and expertise necessary to:
1) carry out individual targeted CRISPR knock outs in mammalian cell lines
- Constitutive CRISPR KO in mammalian cell line using Lentiviral vector systems
- Inducible CRISPR KO in mammalian cell line using Lentiviral vector systems
2) utilize genome wide and targeted CRISPR screening approaches in cancer research including both in vitro and in vivo screens.
- CRISPR screening project consultation to assist investigators in study design.
- Provide lentivirus packaged stocks of several standard genome wide and targeted CRISPR libraries for human, and mouse studies including CRISPR KO and CRISPRa libraries (see stock list below for details)
- Provide lentivirus packaged stocks of custom CRISPR libraries (see stock list below for details)
- Optimize transduction of investigator provided mammalian cell lines with individual CRISPR libraries for CRISPR screening projects
- Provide pooled mutant cell lines from library transduced mammalian cell lines to investigators for carrying out in vitro and in vivo CRISPR screens.
Screen ready stocks of CRISPR libraries
CFS prepares screen ready genome wide libraries for the most commonly utilized genome wide CRISPR libraries as well as selected cancer relevant targeted CRISPR libraries. While the focus is on human CRISPR libraries, several non-human resources have been developed and are available for use. Similarly, the focus of the facility is the development of lentivirus-based screening libraries, additional vectors, such as AAV, can be utilized based on investigator requirements.
Custom CRISPR libraries
The facility can construct targeted CRISPR KO libraries for project driven goals of individual research teams in appropriate vectors. Previously generated custom libraries which are also available for general use can be found here (link to a list we will generate).
The Functional Screening facility works closely with the Cancer Center’s Biostatistics & Computational Biology Shared Resource group to provide necessary computational support for effective utilization of CRISPR screens.