The Center for Advanced Genome Engineering (CAGE) provides expertise and infrastructure for St. Jude investigators to educate, assist, expedite and perform established and emerging genome engineering technologies, including the CRISPR-Cas9 technology. The CAGE has 3 main objectives:
- Introduce user-defined missense mutations, knock-out, and knock-in mutations in human, mouse, rat and Drosophila cell lines as well as human pluripotent stem cells.
- Design and generate the materials and validated genotyping protocols to streamline the rapid production of knockout and knock-in preclinical animal models in collaboration with the existing St. Jude Transgenic/Gene Knockout Facility.
- Identify, validate and introduce newly emerging genome editing technologies and applications to the St. Jude community.
OVERVIEW
The CAGE is a world-class genome editing facility with the mission to provide low cost, expeditious, and collaborative genome engineering services to St. Jude investigators. The CAGE boasts the expertise and infrastructure to educate, assist, expedite, and perform established and emerging technologies, including CRISPR/Cas9.
IMPACT
The genome engineering field is rapidly evolving. Accordingly, the CAGE spends time identifying, testing, and evaluating the latest genome editing tools, technologies, and applications. CAGE has worked with over 75 St. Jude investigators across 15 departments. Since inception in 2017, CAGE scientists have enabled, collaborated on, or published over 50 research papers in high quality journals including Cell, Nature, Nature Communications, Nature Genetics, and Nature Neuroscience. Additionally, team members have been collaborating authors on 29 publications in high quality journals.
“Before I was even on payroll I started developing CRISPR lines…and by the time I landed in January, those lines were in a freezer and [CAGE] was saying, ‘OK, what’s next?’ No other place is going to afford you that.”
Andrew Kodani, Assistant Member
Services
- Introduce user-defined knockout and knock-in mutations in human, mouse, and rat cell lines including in human pluripotent stem cells.
- Design and generate the reagents and validated genotyping protocols to streamline the rapid production of knockout and knock-in animal models in collaboration with the existing St. Jude Transgenic Mouse Facility.
- Facilitate pooled gRNA screening by designing, assembling, and validating pooled gRNA libraries as well as assisting in downstream NGS sequencing and analysis after a screen has been completed.
- Identify, validate, and introduce newly emerging genome editing technologies and applications to the St. Jude community.
- The CAGE recently launched a new Cell Fitness/Dependency Assay which allows investigators to rapidly follow up on pooled knockout screens and assess if a cell line is dependent on a given gene
Technology and advanced equipment
- (4) Tecan automated liquid handler robots
- Mi-seq next-gen DNA sequencer
- (2) semi-automated benchtop pipettors
- (2) Multidrop Combi dispensers
- Lonza nucleofector 4D X-Unit
- Lonza nucleofector shuttle
- EVOS fluorescence imaging system
Dr. Shondra Pruett-Miller
Dr. Shondra Pruett-Miller is the director of the Center for Advanced Genome Engineering (CAGE) at St. Jude. She earned her PhD in Cell and Molecular Biology from UT Southwestern Medical Center and was the founder and director of the Genome Engineering and iPSC Center (GeiC) at Washington University in St. Louis before founding CAGE shared resource at St. Jude in 2017. Her interest in genome-editing technologies, disease modeling in iPSCs, and development of preclinical models using CRISPR-Cas9 help shape the extensive resources that CAGE provides investigators at St. Jude.
