The Genome Engineering team assists Stowers Institute researchers in the precise and specific alteration of genetic material of research organisms and cultured cells.
The Genome Engineering team provides expertise in CRISPR technology and its application in several model systems, including mouse, zebrafish, cavefish, fruit fly, sea anemone, apple snail, and cultured cell lines. Projects begin with the precise design and preparation of custom guide RNAs and homology-directed repair donor templates. These molecular tools are then electroporated into cultured cells to generate engineered lines. For animal model projects, the team prepares the custom reagents and delivers them into embryos.
To identify the engineered mutations, the team provides a targeted deep sequencing service. This allows quick identification of successfully mutated samples and enables the team to process hundreds of animal tissue and cell samples each week.
The Genome Engineering team also provides support for pooled screens. These services encompass genome-wide CRISPR screens, nanobody screens, site-saturation mutagenesis, or novel pooled screens. Services include building or acquiring new pooled libraries and the preparation and validation of libraries by next-generation sequencing. Following a screen, the resulting cell populations are processed by the team for genomic DNA purification, target amplification, and next-generation sequencing.
Team Contact
Head, Genome Engineering
Stowers Institute for Medical Research
In 2024 Hassebroek was named Head of Genome Engineering where she works with a team and researchers to generate CRISPR modified cell lines, zebrafish, mice, and many more organisms.
The Genome Engineering team manages projects to generate knockouts, large insertions and deletions, and point mutations in various model organisms. The team customizes each project to produce it as efficiently as possible and use the latest CRISPR technology methods.
Additional services are offered for pooled screens, TAIL-PCR.
Pooled screen support includes building and validating pooled libraries and processing samples resulting from pooled screens.
The TAIL-PCR service locates random integration events in genomic DNA.
All genome engineering projects start with a targeted double-strand break and the design of a guideRNA. For large insertion projects that require a homology-directed repair donor template, the team designs and produces long single-stranded DNA.
These services include cell line modification using custom CRISPR reagents. This comprises treating the cell line, maintenance of cells, and single-cell clone identification by working with Cells, Tissues, and Organoids. The Genome Engineering team prepares custom CRISPR reagents for delivery to animal embryos.
Each week the team processes hundreds of samples from cells and animal tissues to detect and identify the precisely engineered mutations. The powerful workflow the team has built around targeted deep sequencing enables the successful completion of genome engineering projects.
A pooled screen is a powerful way to look across a genome or a massive number of sequences to identify important genes of interest, effects on expression, or significant sequence motifs.
The Genome Engineering team acquires, prepares, and validates pooled libraries as an Institute resource. To coordinate the total pooled screen workflow, the team collaborates with other teams to track the progress of these projects. Once resulting cell populations from a screen are delivered, the team does genomic DNA purification, target amplification, and preparation for NGS.
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A Q&A with Scientific Project Manager Victoria Hassebroek
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Anoja Perera is the Director of Sequencing and Discovery Genomics at the Institute. She shares more about how her role at the institute supports our labs and scientific research.
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