Methods of constructing a gene mutation library and compounds and compositions thereof

A method of selecting a cell line or a non-human transgenic animal model for the analysis of the function a gene comprising:

• a) introducing into an embryonic stem cell a vector having a selectable marker which, when the vector is inserted within the gene, the inserted vector can inhibit the expression of the gene;
• b) selecting embryonic stem cells expressing the selectable marker;
• c) excising the vector from the embryonic stem cells expressing the selectable marker whereby host DNA from the gene is linked to the excised vector;
• d) sequencing host DNA in the excised vector;
• e) comparing the sequence of the host DNA to known gene sequences to determine which host DNA is from a gene for which a model for the analysis of the function the gene is desired;
• and
• f) selecting the embryonic stem cell containing the inhibited gene for which a model for the analysis of gene function is desired.

A method of creating a library of embryonic stem cells wherein 1) a multiplicity of cells in the library each contain a gene having inhibited expression, 2) a sequence of the gene having inhibited expression is known, and 3) a multiplicity of different inhibited genes is represented in the library, comprising

• a) introducing into an embryonic stem cell a vector having a selectable marker which, when the vector is inserted within a gene, the inserted vector can inhibit the expression of the gene;
• b) selecting embryonic stem cells expressing the selectable marker;
• c) excising the vector from the embryonic stem cells expressing the selectable marker such that host DNA from the gene is linked to the excised vector;
• and
• d) sequencing the host DNA in the excised vector;
• thereby identifying sequence of the gene whose expression is inhibited, and creating a library of embryonic stem cells containing the gene whose expression is inhibited and a sequence of the inhibited gene is known.