An evaluation of techniques to enhance incorporation of foreign DNA into the mammalian genome /
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| Other Authors: | , , |
| Format: | Thesis Book |
| Language: | English |
| Published: |
1989.
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| Subjects: | |
| Online Access: | Link to OAKTrust copy |
| Abstract: | The objectives were to evaluate the use of potential DNA damaging agents (DAPI stain and UV radiation) to enhance transgene incorporation into mouse zygotes, and to evaluate phospholipid vesicles (liposomes) for the delivery of DNA to uterine stage mouse and bovine embryonic cells. Data set 1) 762 mouse zygotes were injected (1 pg/ul pMK) across 4 treatments representing UV radiation, DAPI staining, DAPI/UV in combination, and controls (A,B,C,D). Pregnancy rates were 40.0%, 40.0%, 10.0%, and 20.0% with percent offspring survival of 9.5%, 5.3%, 1.1%, and 3.2%, respectively. No transgenic mice were produced. No significant differences were detected between groups (P>.05). Data set 2) 850 zygotes were injected (1 ng/ul pMK) and pregnancy rates across treatments were 35.7%, 41.7%, 0.0%, and 53.3% with percent offspring survival of 11.4%, 9.4%, 0.0%, and 11.6%, respectively, with two transgenic animals produced in treatment B (11.8% of offspring) and 3 transgenics in treatment D (11.5% of offspring). No differences were detected between groups (P>.05). The treatments did not enhance transgene incorporation. Liposome-encapsulated DNA (L-DNA) or DNA in buffer was injected into mouse blastocysts. Pregnancy rates were 64.7% and 70.6%, with percent offspring survival of 32.9% and 31.2%, respectively. No transgenic mice were detected and no differences were detected between groups {P >.01). L-DNA was injected into 58 bovine blastocysts (fresh, cryopreservation after injection, and injection after cryopreservation) resulting in pregnancy rates of 36.8%, 66.7%, and 40.0% with fetal survival (40 to 60 days of age) of 28.9%, 30.0%, and 30.0%, respectively. No transgenic fetuses were detected. Delivery of DNA to the embryonic cells via liposomes did not occur at the efficiency needed to detect a transgenic animal with the number of observations used. |
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| Item Description: | Typescript (photocopy). Vita. "Major subject: Veterinary Physiology." |
| Physical Description: | ix, 151 leaves : illustrations ; 29 cm |
| Bibliography: | Includes bibliographical references. |