The recent and unanticipated successes in both cloning sheep using nuclei from adult fibroblasts and in solving many forms of infertility by ICSI, now raises questions as to whether a combination of these approaches will result in reliable and efficient methods for propagating valuable research models, especially NHPs. The technologies routinely available for creating invaluable rodent models for various diseases and many serious human disorders are sometimes not appropriately studied in these lower mammals. Furthermore, spontaneous mutations resulting in unique animals to investigate diseases (e.g., retinitis pigmentosa) are periodically discovered in NHP colonies, and the ability to propagate these animals is vital for future investigations. For these reasons, there are strong justification for developing reliable and effective methods for propagating identical NHPs. While electrofusion of blastomere nuclei with enucleated rhesus oocytes resulted in two offspring (one male, the other female) in 1996, last year's attempts have not been successful. ICNI holds bright promise as an alternative approach for propagating identical NHPs. To that end, this application seeks to sequentially perfect ICNI capabilities and develop new animal resources by addressing twenty-three questions in four Specific Aims: 1) Is ICSI an effective and efficient method for propagating rhesus monkeys?; 2) Is ROSI an effective method for propagation?; 3) Is blastomere ICNI, using blastomere nuclei obtained from rhesus monkey embryos injected into unfertilized enucleated oocytes, an effective and efficient method of propagation?; and 4) Is somatic ICNI, using nuclei obtained from somatic cells cultured from adult and fetal rhesus tissues and injected into unfertilized enucleated oocytes, an effective and efficient method of propagation? By expanding the manner in which reproduction is achieved, the utility of this model for propagating important research animals will be greatly enhanced. ICNI may well find uses as a reliable, routine approach for propagating invaluable research animal resources for the entire biomedical research community.
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