The Animal Modeling Core (AMC) not only applies established genome-engineering technologies but also develops and optimizes new approaches to address challenging animal-model generation projects.
Our team works to improve the efficiency, flexibility, and precision of genome editing by developing novel strategies for targeted DNA integration, allele-specific engineering, embryo manipulation, and other applications. These technologies are designed to expand the types of genetically engineered models that can be generated and to provide solutions when conventional approaches are inefficient or unsuitable.
AAV-Mediated Donor Delivery with 2-Cell Embryo CRISPR Electroporation
The AMC developed a genome-editing strategy that combines adeno-associated virus (AAV)-mediated delivery of DNA repair templates with CRISPR-Cas9 ribonucleoprotein electroporation of 2-cell embryos.
This approach was developed to facilitate efficient targeted DNA knock-in and can be particularly useful for insertions that are difficult to introduce using conventional embryo electroporation methods. The strategy has been successfully applied to both mouse and rat embryo genome editing.
Learn more: Efficient DNA knock-in using AAV-mediated delivery with 2-cell embryo CRISPR-Cas9 electroporation — Frontiers in Genome Editing.
A detailed experimental protocol for application of this technology has also been published in the Journal of Visualized Experiments (JoVE).
CRISPR-SWITCH: Controlled Monoallelic Genome Editing
The AMC developed CRISPR-SWITCH (Silent mutations With Intention To Create Heterozygotes), a genome-engineering strategy designed to enable controlled monoallelic CRISPR editing while preserving an unmodified wildtype allele.
This approach can be particularly valuable when modeling dominant-negative or dosage-sensitive disorders in which conventional CRISPR editing may produce unwanted biallelic modification or homozygous alleles that are lethal or biologically inappropriate.
CRISPR-SWITCH was developed and demonstrated during generation of a heterozygous Syt1-D365E mouse model of Baker–Gordon syndrome.
Learn more: CRISPR-SWITCH: a strategy for monoallelic genome editing and generation of a Syt1-D365E mouse model of Baker–Gordon syndrome — Frontiers in Genome Editing.
Developing New Solutions for Challenging Models
Many animal-modeling projects cannot be addressed with a single standardized genome-editing workflow. The AMC routinely evaluates and develops customized strategies for projects involving difficult genomic loci, complex knock-ins, allele-specific modifications, large DNA insertions, or other technically challenging applications.
Investigators with challenging or unconventional model-development projects are encouraged to contact the AMC to discuss potential approaches.