transgenic mice

Changed or foreign genes can enter mouse embryo cells {transgenic mice} at chromosomal positions. Transgenic-mice descendants have changed or foreign genes and have new proteins.

organism

Mammals have cell and tissue interactions, so testing requires whole organisms.

process: injection

SV40, Moloney murine leukemia virus (MoMLV), or mouse mammary tumor virus (MMTV) microinjection can put changed or foreign DNA into cells. Cloned-gene microinjection into fertilized egg pronuclei can put changed or foreign DNA into cells.

process: cell addition

Mice embryos can change by adding altered cells. Mouse blastocysts have inner-cell {embryonic stem cell, blastocyst} (ES cell) layers, which can culture with fibroblasts or with leukemia inhibiting factor to prevent further differentiation. Embryonic stem cells can uptake and insert genes by homologous recombination. Then ES cells go into mouse embryos.

marker

Neo gene resists G418. ES cells with neo gene resist G418 and live.

embryonic development

In embryos, tissue-specific regulators express changed or foreign genes in one tissue but not different tissues. If changed or foreign genes are toxic, they destroy tissue {cell ablation, toxin}. Ablated cells prevent subsequent tissue development, allowing embryo location and function tracking {cell lineage study}. Retrovirus with E. coli lacZ reporter can trace tissue differentiation and cell migration.

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Biological Sciences>Genetics>Recombinant DNA>Cloning>Transformation

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Date Modified: 2022.0224