The Animal Modeling Core (AMC) provides a broad range of molecular genetics, genome-engineering, and model-support services that complement our mouse and rat model-generation programs. Services can be provided as part of a complete animal-modeling project or as stand-alone support for investigators performing their own in vitro or in vivostudies.
Our team has extensive experience in CRISPR reagent design, molecular cloning, viral vector design, RNA-targeting approaches, genotyping, molecular characterization, and development of customized experimental strategies.
CRISPR-Cas Reagent Design and Validation
The AMC provides design and testing services for CRISPR-Cas genome-editing reagents for animal and cell-based applications.
Services may include:
- CRISPR guide RNA design and sequence analysis
Evaluation of predicted off-target sites
Design of single-stranded and double-stranded DNA repair templates
Design of point mutations, knock-ins, deletions, and other targeted modifications
In vitro testing of CRISPR reagents when appropriate
PCR and sequencing-based evaluation of genome-editing activity
Design and optimization of assays for detecting desired and unintended editing outcomes
Repair templates can include synthetic single-stranded oligonucleotides, long single-stranded DNA, plasmid-based donors, and AAV-compatible donor constructs.
AAV and Expression Vector Design
The AMC can assist investigators with the design and development of customized adeno-associated virus (AAV) expression constructs for in vitro and in vivo research applications.
Vector designs can be customized based on the intended target tissue, experimental application, and desired pattern of gene expression. Applications may include:
Wildtype or mutant gene overexpression
Expression of disease-associated variants
Reporter gene expression
Gene replacement strategies
Knockdown-and-replacement strategies
shRNA or miRNA-based gene silencing
CRISPR and other genome-editing applications
Cell- or tissue-specific expression
AAV-delivered DNA repair templates for genome editing
The AMC can assist with selection and organization of vector components including promoters, coding sequences, reporters, P2A or other multicistronic elements, regulatory sequences, and polyadenylation signals while accounting for AAV packaging constraints.
Depending on project needs, services can include vector design, plasmid construction, sequence verification, and assistance developing an appropriate experimental strategy for subsequent AAV production and use.
RNA-Targeting and Oligonucleotide Design
The AMC provides design support for RNA-targeting approaches that can be used for gene knockdown, allele-specific targeting, therapeutic development, and proof-of-concept studies.
Services may include:
Short interfering RNA (siRNA) design
Allele-specific siRNA design
Short hairpin RNA (shRNA) and micro-RNA (miRNA)-based knockdown construct design
Antisense oligonucleotide (ASO) design
Design of oligonucleotides targeting splice sites or other RNA-processing events
Sequence analysis to identify candidate target regions
Evaluation of specificity and potential off-target sequences
Design of appropriate molecular assays to evaluate target knockdown or transcript modification
For projects involving disease-associated variants, the AMC can also assist with development of strategies designed to preferentially target mutant transcripts while preserving expression of the wildtype allele.
RNA-targeting services can be integrated with cell-based testing, AAV vector development, or animal-model studies depending on project needs.
Molecular Genetics and Custom Cloning
The AMC offers customized molecular biology and cloning services to support animal-modeling, genome-editing, and related research projects.
Services may include:
Custom plasmid design and construction
Expression vector construction
CRISPR targeting vector and repair-template construction
Reporter and fusion-protein constructs
Construction of genotyping and qPCR standards
Sequence verification and construct characterization
Projects requiring specialized or complex cloning strategies are evaluated individually.
Genotyping and Molecular Characterization
The AMC provides custom genotyping assay development and molecular analysis for genetically engineered animal models and other experimental samples.
Services may include:
Custom PCR primer and assay design
DNA extraction from animal tissues or other samples
Endpoint PCR
Restriction fragment length polymorphism (RFLP) analysis
High-resolution melt (HRM) analysis
Sanger sequencing-based genotype confirmation
Sequence analysis of genome-edited alleles
Droplet digital PCR (ddPCR)
Transgene or allele copy-number analysis
Evaluation of founder animals with potentially complex genome-editing outcomes
For newly generated models, the AMC can develop customized strategies to distinguish wildtype, heterozygous, homozygous, conditional, and other genetically modified alleles.
Colony Management
The AMC provides organized management of mouse and rat breeding colonies for both internal and external investigators.
Services may include:
Establishment and maintenance of breeding colonies
Breeding strategy development
Setting up and monitoring matings
Weaning and animal identification
Tissue collection for genotyping
Genotype tracking
Producing and shipping experimental cohorts
Colony record keeping
Breeding strategies can be customized to efficiently generate experimental and control animals with the desired genotypes.
Model Characterization
The AMC can assist investigators with molecular and phenotypic characterization of newly generated animal models.
Characterization strategies are customized based on the model and research question and may include:
DNA-level confirmation of the engineered allele
RNA expression analysis
Tissue collection and processing
Imaging
Histological analysis
Coordination or development of specialized phenotyping assays
The AMC can also work with investigators and other University of Missouri core facilities to develop an appropriate characterization strategy when specialized expertise or instrumentation is required.
Custom Projects and Experimental Design
Many projects do not fit within a standard service category. AMC investigators routinely collaborate with researchers to develop customized approaches involving molecular genetics, genome editing, animal modeling, viral vectors, RNA-targeting technologies, and related applications.
Investigators are encouraged to contact the AMC early in project development. Our team can help evaluate feasibility, compare experimental approaches, develop project workflows, and identify the most appropriate technologies for addressing a research question.
AMC faculty are also available to collaborate on grant applications and research projects that would benefit from expertise in genome engineering, molecular genetics, and animal-model development.