Question

Review of Concepts: Gene Identification and Gene Knockout You want to study the physiological role of a gene you just cloned by making a "Knock-out" mutant mouse. Your gene has 5 exons (see below) and you want to mutate it by inserting neomycin resistance gene (neo) into exon 3 1 2 probe 3 4 Eco RI Eco R1 Eco Ri

2) You think that the neomycin resistance gene (neo) is causing an artificial result, so you also make a “knock-out” mutant using the LoxP-Cre system to splice-out the neo gene. Briefly, describe how you would accomplish this task. Draw a picture of the targeting vectors and a map of the gene before and after the neo gene has been removed

Answer 1

. Cre-lox recombination for the removal q neo gene L2 Etheo = - X 3 – luxe loxP Promoter . cre recombinese scoe recombincese t y cheesy - nico gene Spliced out. 04244357

cre - loxP system permits development of recombinant mice in which gene is knocked out in a specific cell/tissue. In the process, A lox P site is inserted on each side of neo exon of the traget gener by homologous recombination. The Cre gone, which produces recombincase enzyme will be expressed on opposite call type by non- homologous recombination. Crossing of both cells lead to the delehu. of specife exon (heo) by are protein.