Texas A&M Institute for Genomic Medicine

The Texas A&M Institute for Genomic Medicine (TIGM) is an essential resource for any researcher looking to obtain knockout mice and embryonic stem (ES) cells quickly and with favorable intellectual property (IP) terms. Our resources include the world’s largest gene trap library of ES cells in the C57BL/6N mouse strain and a constantly expanding repository of cryopreserved germplasm of knockout lines. TIGM provides both ES cell clones and mice as well as other transgenic core services including CRISPR/Cas9-based genome modifications within the Texas A&M system and to the public and private international research community.

Mouse Transgenic Services

Basic Pronuclear/Cytoplasmic Injection (including CRISPR/Cas9*): Investigator provides at least 30 ug of digested plasmid DNA, detailed map of the construct and a picture of the DNA with the desired band circled. In case of CRISPR, investigator provides uncut plasmid vector expressing sgRNA, sequences of the sgRNA and template DNA vector of oligo as needed. Includes injection of at least 125 FVB embryos. All resulting offspring will be transferred to the investigator at weaning. TIGM can also genotype the offspring for the transgene at an extra cost.

Pronuclear Injection, other strains (C57BL/6, etc.)

Pronuclear Injection of BAC DNA: Please contact the core to obtain protocols for preparing BAC DNA. BAC DNA needs to be submitted to the core as a pellet under 70% ethanol.

Blastocyst Injection: Investigator provides two vials of the frozen clone. The service includes injection of at least 30 blastocysts/clone. All resulting offspring will be transferred to the investigator at weaning.

Sperm Cryopreservation: Sperm from two male mice will be cryopreserved using the protocol from the Jackson Laboratory. The cryopreservation fee includes sperm viability testing by in vitro fertilization.

Sperm Cryopreservation (Jax kit): Sperm from two male mice will be cryopreserved using the kit from the Jackson Laboratory. The viability of the frozen sperm will be tested at the Jackson Laboratory by in vitro fertilization and stored at the Jackson Laboratory.

Embryo Cryopreservation: Investigator transfers 6-8 practiced stud males, 3-6 months of age, to TIGM quarantine. The core will perform two sessions and will cryopreserve at least 200 embryos at the morula stage.

Rederivation via IVF (live mice): Investigator transfers 2-3 practiced stud males, 3-6 months of age, to TIGM quarantine for sperm collection. The core will isolate oocytes from at least 5 superovulated females (C57BL/6, FVB or CRD1 ICR strains; other strains could be used too, but additional charges related to animal order may apply), perform IVF and transfer the resulting embryos to recipient females. All resulting offspring will be transferred to the investigator at weaning. The remaining sperm sample can be cryopreserved and stored at TIGM.

Rederivation via IVF (sperm samples prepared using Jax or more recent protocol): Investigator provides at least 2 straws each containing at least 40-50 ul of frozen sperm. The core will isolate oocytes from at least 5 superovulated females (C57BL/6, FVB or CRD1 ICR strains; other strains could be used too, but additional charges related to animal order may apply), perform IVF and transfer the resulting embryos to recipient females. All resulting offspring will be transferred to the investigator at weaning.

• Rederivation via IVF (sperm samples prepared using other protocols): Investigator provides at least 4 straws/vials each containing approximately 200 ul of frozen sperm. The core will perform two sessions and will use oocytes from 20 donors per vial of sperm. C57BL/6, FVB or CRD1 ICR strains could be used as donors; other strains could be used too, but additional charges related to animal order may apply. The remaining straws will be kept in reserve for another transfer attempt if necessary. All resulting offspring will be transferred to the investigator at weaning.
• Rederivation via Embryo Transfer: Investigator transfers 3-6 practiced stud males, 3-6 months of age, to TIGM quarantine for mating with superovulated females. The core will perform 1-2 sessions and will transfer at least 100 embryos at the morula stage.
• Frozen Embryo Transfer: Investigator provides at least 4 straws each containing approximately 20 embryos. 2 straws will be used for transfer. The remaining straws will be kept in reserve for another transfer attempt if necessary. All resulting offspring will be transferred to the investigator at weaning.
• Live Embryo Transfer: Investigator provides early stage embryos in M2 medium or alternatively freshly isolated uterine or oviduct tissues from timed pregnancies for the core to flush. The investigator must coordinate with the core at least a week in advance so that the core can set up recipient females at the appropriate stage. All resulting offspring will be transferred to the investigator at weaning.
• Colony maintenance: Includes general husbandry, health monitoring, breeding, weaning, ear tagging, tail clipping and genotyping as needed.
• Gene Targeting via Homologous Recombination (constitutive, conditional or knock-in) Services include:
• Targeting vector construction and designing of the screening strategy
• Transfection of ES cells and confirmation of targeted clones
• ES cell blastocyst injection
• Chimera Breeding
• Genotyping protocol design and confirmation
• Electroporation service package: The service includes linearization/purification/ electroporation of targeting construct; isolation/expansion of drug-resistant colonies in 96-well format (at least 2 plates); preparation of cell pellet plates in 96-well format and expansion of all targeted ES cell clones as specified by the customer.

Additional services:

• Identification of targeted ES cell clones by PCR
• Expansion of targeted ES cell clones

Equipment:

Imaging

• ultra-high resolution imaging and dual-energy x-ray absorptiometry (Faxitron Ultra Focus Digital Radiography)
• high-resolution X-ray CT (Scanco Viva40 MicroCT)
• fluorescence (Bruker In-Vivo Xtreme, Xenogen – IVIS ® Lumina II, NightOWL)
• luminescence (Bruker In-Vivo Xtreme, Xenogen – IVIS ® Lumina II, NightOWL)
• radioisotopic (Bruker In-Vivo Xtreme)
• high resolution X-ray (Bruker In-Vivo Xtreme)

Physiology systems

• DEXA for bone density analysis

Pathology

• Gross Observations
• Organ Tissue Weights
• Histopathology

Expression studies

• Taqman
• Microarrays