Transfer of genes to fertilized eggs or embryos:The transfection of fertilized egg includes either the transfer of entire nuclei or entire chromosomes; or their fragments or the DNA segments.
A. For the transfer of entire nuclei, the cells are treated with cytochalasin B and enucleated. The enucleated cells are hatched with the desired karyoplasts (i.e., nuclei) for induction in the presence of polyethylene glycol (i.e., PEG).
B. For transfer of entire chromosomes, metaphase cells are subjected to hypotonic lysis and individual fragments or chromosomes are isolated and then incubated with entire cells or eggs for transfection.
C. Microinjection of DNA segments: In this the fertilized eggs are injected with DNA segments for the integration. DNA integrated eggs are then employed for acquiring transgenic animals.
D. Transfer of genes to cultured cells: In this stem cells are utilized. The stem cells are undifferentiated precursor cells. In such cultured cells, the gene can be delivered via vectors such as retroviruses or directly by methods like microinjection employing particle gun, electroporation or by the utilization of liposomes.
Transgenic animals have been generated in a variety of animals like rabbits, mice, sheep, goats, pigs, cows, fishes and so on.
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