Question

Which of the following are steps in the generation of knockout mice using ES cells?

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• Introduce chimeric host blastocyst into a surrogate mother mouse.Introduce chimeric host blastocyst into a surrogate mother mouse.

• Cross chimeric mouse to a wild-type mouse to obtain a heterozygous knockout mouse.Cross chimeric mouse to a wild-type mouse to obtain a heterozygous knockout mouse.

• Make a construct with genomic DNA corresponding to the ends of the gene to be knocked out on either end of a drug resistance marker gene.Make a construct with genomic DNA corresponding to the ends of the gene to be knocked out on either end of a drug resistance marker gene.

• Introduce heterozygous knockout ES cells to a host blastocyst.Introduce heterozygous knockout ES cells to a host blastocyst.

• Introduce a gene-targeting construct to ES cells growing in a petri dish.Introduce a gene-targeting construct to ES cells growing in a petri dish.

• Apply selection for drug resistance marker.Apply selection for drug resistance marker.

• Cross two heterozygous knockout mice to each other to generate a homozygous knockout.Cross two heterozygous knockout mice to each other to generate a homozygous knockout.

• Add LTRs to the ends of a cDNA for the gene to be knocked out.Add LTRs to the ends of a cDNA for the gene to be knocked out.

• Introduce a gene-targeting construct into host blastocysts.Introduce a gene-targeting construct into host blastocysts.

• Mix modified ES cells with ES cells containing a Cre gene.Mix modified ES cells with ES cells containing a Cre gene.

• Introduce a gene-targeting construct into packaging cells.Introduce a gene-targeting construct into packaging cells.

• Replace the viral genes with a therapeutic gene.

Answer 1

Answerall of the steps are neccesary:

• All the steps from 1 to 12 are necessary to produce the knock out mice.
• The first knock out mice was prepared by Capeecchi, Martin and Smithies in the year 1989, and they were awarded Nobel Prize for it in the year 2007.
• For preparing the knock out mice, the original genes of the mice are knocked out and the gene of interest is inserted.
• After following all the steps the homozygous knock out mice is prepared.
• They are quite useful for the experimentation on various physiological processes in the human beings, because our general physiology matches to the mice.
• Such type of mice can be helpful for many amateur discoveries

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