SBIR-STTR Award

The ability to genetically modify mice through methods of gene transfer in fertilized mouse embryos (transgenic mice) or gene modification in embryonic stem (ES) cells (gene targeting) has dramatically increased the value and use of mice in biomedical research. . The production of gene-modified mice requires the surgical transfer of embryos into female mice. This surgery requires anesthesia (with potential complications), post-operative care of female mice (which may include the use of analgesics), as well as technical expertise on the part of the person carrying out the surgery, as. At the same time, there is a strong desire in the animal research community to reduce the morbidity, stress and pain of animals used for research purposes. To overcome these problems, we have developed a Non-Surgical Embryo Transfer (NSET) device that can be used for the non-surgical transfer of embryos into female mice. This procedure is rapid (transfer requires about 10 seconds), does not require surgery (including the associated anesthesia and post-operative care), and requires little expertise. Our preliminary data indicate that our NSET device is equally successful as standard surgical embryo transfer in mice in the production of transgenic mice. Preliminary studies were performed with hand-made NSET devices. We have now manufactured a NSET device, enabling us to produce large numbers of NSET devices with a high degree of uniformity. The aims of this proposal are to demonstrate the efficacy of this manufactured NSET device for the non-surgical transfer of embryos that (1) have may be transgenic by DNA microinjection, (2) have been cryopreserved, and (3) were generated by in vitro fertilization of embryos with cryopreserved sperm. Success in these aims will provide the foundation for the commercialization of the NSET device for academic and bio/pharma mouse facilities.

Public Health Relevance:
We have developed a device for the non-surgical transfer of embryos into recipient female mice that represents a substantial advance over existing methods of generating gene-modified mice in that it eliminates surgery, including anesthesia and post-operative care. The goal of this proposal is to determine the efficacy of a manufactured non-surgical embryo transfer device for the transfer of embryos that are transgenic, cryopreserved, or generated by in vitro fertilization.

Public Health Relevance Statement:
Narrative We have developed a device for the non-surgical transfer of embryos into recipient female mice that represents a substantial advance over existing methods of generating gene-modified mice in that it eliminates surgery, including anesthesia and post-operative care. The goal of this proposal is to determine the efficacy of a manufactured non-surgical embryo transfer device for the transfer of embryos that are transgenic, cryopreserved, or generated by in vitro fertilization.

Project Terms:
Analgesic Agents; Analgesic Drugs; Analgesic Preparation; Analgesics; Anesthesia; Anesthesia procedures; Animal Experimental Use; Animal Experimentation; Animal Research; Anodynes; Antinociceptive Agents; Antinociceptive Drugs; Biomedical Research; Caring; Communities; Contracting Opportunities; Contracts; DNA; Data; Deoxyribonucleic Acid; Development; Devices; ES cell; Embryo; Embryo Transfer; Embryonic; Expertise, Technical; Female; Fertilization in Vitro; Foundations; Fungi, Filamentous; Gene Targeting; Gene Transfer; Gene-Modified; Genes; Goals; Grant; Hand; Hand functions; IVF; Lead; Legal patent; Licensing; Mammals, Mice; Mediating; Medical; Methods; Mice; Mice, Transgenic; Microinjections; Modification; Molds; Morbidity; Morbidity - disease rate; Mouse Strains; Murine; Mus; Operation; Operative Procedures; Operative Surgical Procedures; Pain; Painful; Patents; Pb element; Performance; Persons; Phase; Post-Operative; Postoperative; Postoperative Period; Procedures; Production; Research; Savings; Scientist; Speed; Speed (motion); Sperm; Spermatozoa; Stress; Surgical; Surgical Interventions; Surgical Procedure; Targetings, Gene; Technical Expertise; Technology; Test-Tube Fertilization; Time; Transgenic Mice; Transgenic Organisms; Using hands; Work; analgesia; animal pain; base; commercialization; cost; embryo transplantation; embryonic stem cell; heavy metal Pb; heavy metal lead; improved; large scale production; novel; public health relevance; sperm cell; stem cell of embryonic origin; success; surgery; transfer of a gene; transgenic; zoosperm

The use of research animals in research has been essential to development of vaccines and study of most human diseases. Modern rodent research is increasingly powerful with the ability to manipulate the genomes of mice and rats such that they closely mimic complex human diseases such as Alzheimer's and artherosclerosis. Generating transgenic mice and rats, requires that embryos that are manipulated by researchers be transferred into recipient female mice where they can complete their development. Up to this point, these transfers have required surgical procedures in which the embryo is implanted into the uterus. Recently, a device has been developed that enables these embryo transfers to be performed without surgery which eliminates the post-operative recovery period thereby reducing pain of the animals. This proposal investigates and expands upon the utility of this embryo transfer device by establishing the optimal ages and mouse strains for embryo transfer (Aim 1), determining whether embryonic stem cells can be effectively and efficiently transferred with this device (Aim 2), determining whether the device is useful for artificial inseminations (Aim 3), directly visualizing the device in the uterus which may enable design improvements (Aim 4) and quantifying measures of stress relative to surgical procedures to support widespread adoption of this procedure in the research community (Aim 5). The primary project objective is to produce data that will prove the efficacy of this embryo transfer device and thereby enable researchers to replace surgical procedures with a non-surgical method that is equally effective. Proving that this non-surgical transfer method is effective under a range of conditions will expand its use over this range of applications and thereby maximize its impact in enabling researchers to refine and reduce the number of surgical procedures performed in mice.

Public Health Relevance:
Animal use in research is essential for biomedical research and with the developing abilities to genetically engineer mice for research;these animals are becoming even more important as they can be modified to mimic most human diseases. Genetic engineering of mice requires, as one of many other things, the ability to transfer embryos into mice where they complete their development. This project supports further development of a device that enables these embryo transfers to be performed without surgery and therefore greatly reduces the pain category that these research animals experience and thereby supports progress in biomedicine.

Thesaurus Terms:
Address;Adoption;Age;Alzheimer;Alzheimer Disease;Alzheimer Sclerosis;Alzheimer Syndrome;Alzheimer's;Alzheimer's Disease;Alzheimers Dementia;Alzheimers Disease;Animal Experimental Use;Animal Experimentation;Animal Research;Animals;Artificial Insemination;Biomedical Research;Biotechnology, Genetic Engineering;Blastocyst;Blastocyst Transfer;Blastocyst Structure;Blastosphere;Categories;Collaborations;Common Rat Strains;Communities;Complex;Data;Dementia, Alzheimer Type;Dementia, Primary Senile Degenerative;Dementia, Senile;Development;Device Designs;Devices;Es Cell;Embryo;Embryo Transfer;Embryo, Preimplantation;Embryonic;Ethics;Eutelegenesis;Fallopian Tubes;Female;Fertilization In Vitro;Gene-Modified;Generations;Genetic Engineering;Genetically Engineered Mouse;Genital System, Female, Uterus;Harvest;Ivf;Implant;In Vitro;Investigators;Mammalian Oviducts;Mammals, Mice;Mammals, Rats;Mammals, Rodents;Measures;Mediating;Methods;Mice;Molecular Biology, Genetic Engineering;Mouse Strains;Murine;Mus;Operation;Operative Procedures;Operative Surgical Procedures;Pain;Painful;Performance;Phase;Position;Positioning Attribute;Post-Operative;Postoperative;Postoperative Period;Primary Senile Degenerative Dementia;Procedures;Production;Rat;Rattus;Recombinant Dna Technology;Recovery;Relative;Relative (Related Person);Research;Research Personnel;Researchers;Rodent;Rodentia;Rodentias;Salpinx;Sperm;Spermatozoa;Staging;Stress;Surgical;Surgical Interventions;Surgical Procedure;Technology;Test-Tube Fertilization;Testing;Transgenic Mice;Uterine Tubes;Uterus;Veterinarians;Animal Pain;Biomarker;Blastocyst;Blastula;Community;Cost;Dementia Of The Alzheimer Type;Design;Designing;Egg;Embryo Transplantation;Embryonic Stem Cell;Experience;Human Disease;In Vitro Fertilization;In Vivo;Mouse Genome;Oviduct;Primary Degenerative Dementia;Prospective;Senile Dementia Of The Alzheimer Type;Sperm Cell;Stem Cell Of Embryonic Origin;Success;Surgery;Vaccine Development;Womb;Zoosperm