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 custom design and preparation of guideRNA and homology-directed repair donor templates. Next, the team uses these custom molecular reagents to engineer cultured cell lines directly. For animal model projects, the team prepares the custom reagents and hands them off to the Stowers’ animal technology centers and researchers for embryo delivery.
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.
Genome Engineering now offers a service utilizing TAIL-PCR to identify random integration events. The team has successfully used this method in killifish, sea anemone, fruit fly, and zebrafish.
The team is also responsible for maintaining and distributing 137,000+ clones and vectors. The thirty-four collections of clones include yeast, fly, mouse, human, and expression vectors.