Groups # 10Mouse knock-inMirna ZhouNhan NguyenEfren SandovalWhat is it used for?! In Gene knock-in a coding or regulatory region is Exchanged orAltered:• Change endogenous gene for homologous Exogenous DNA" Do homologous genes among species have the samefunction?• Altered endogenous gene properties: e.i. change codon (aminoacid)" To Study specific gene function• Model for human diseases (In known gene alteration andphenotype)" Study, Understand and Create new & efficient therapiesfor diseases1HOW ISIT DONE?2A gene knock-in construct contains:1.Gene of interest2.Homologous DNA sequences at the 5’and 3’ arms enclosing the gene of interest3.(+/-) selectable markers which would beeither resistance or sensitive to antibiotics.1HOW ISIT DONE?23 Knock-inEmbryonic Stem cell4567ElectrophorationExamples:1. Homologous gene replacement: Pax 5 can substitute for Pax 2 in eye development2. Reporter: Visualizing gene expression using LacZ and GFP under control of endogenous regulatory regionsWild type Pax 5 Mutant replacementHuntington’s disease• HD example is used in helping to determine the as yet unknownmolecular mechanisms underlying neural dysfunction in HD• The late onset neurological abnormalities displayed by theses miceprovide both end stage neuroanatomical features and a set ofsimple non-invasive behavioral assays that will be valuable in theassessment of therapies designed to cure HD• Gene knockin strategies have been used to replace or mutate genesthat affect cardiac electrophysiology in the mouse and to engineermodels of human disease, particularly those potentially associatedwith gain of function
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