What would be the outcome if the procedure used to make gene-targeted (knock-out) mice was altered in the following ways (consider each situation separately).
(a) The embryo-derived stem (ES) cells did not possess a gene responsible for visible phenotypic trait (such as agouti coat color) that differed from that of the cell in the recipient blastocyst.
(b) The inserted "disrupting" gene (e.g. neoR) did not insert into the gene of interest.
(c) Chemical transfection or electroporation failed.
(d) The gene knock-out was embryonic lethal
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