saikrishnaVenkatasubramanian Honglu FlorenceClifford TracyPham JelenaGrabjelsek

1)DNAmethylationrecruitshistoneswhichresultsintherepressionofthegeneexpression 2)Imprintingisaphenomenawhereoneof2alleleissilencedthroughpassageduringgermline production 3)RegionwithmanyregulatoryelementsthatseemtocontroltheexpressionofHoxGenes 4)CreLoxsystem,loxpsiteswereinoppositedirectionswhichresultedinaninversion 5)DigitEnhancerSequencesarewhatthegreenovals 6)TheyusedthetransgeneiclinewiththeHoxd9Laczreportergeneasatransgeneandthe othertransgeniclineiswitha2.7mbinversionofthehoxDinv(rel5itag6) b)DatashowsthatsameLaczexpressioninbothparentaltypes,whereasinthecaseof inversiononlythematernalgenesarecompletelyrepressedandnoexpressionofhoxd9is shownandPaternalexpression c)Eventhoughthegenomeisthesame,therewasadifferentialexpressionbetweenpaternal andmaternaltypes d)Maternalrepressionexpressionwaspositiondependantandtheeffectissexspecificandthe presenceLacZwasrequiredforthematernalrepression,toshowthistheyshowed2lineswith HoxD11withlacZandwithout,andexpressionwasonlydetectedinsituationwithLacztherefore botharerequiredfortheexpression 7)Inversionisspecifictothepositiondependantasthepositionofthereportergenewas altered/movedby2.7mbandthusleadtothedifferentialexpressioninbothmaternalandpaternal casesoftheHoxD9gene. 8a)Theblackcirclesrepresentthatthemethylationisoccurring,whitecirclesshowthatthereis nomethylationisoccurring.androwsofcircleshowseachindividual. b)Thereisnodifferenceinmethylationbetweennoninvertedrel5forindividualswhorecieved maternalorpaternaltransgenes.Whereasthereisisaclearmaternalbiasfortheinverted rel5itag6inversionshowingstrongmethylation.Inthecasesofescaperembryosthereareodd expressionswhereinversionleadstoexpressioninsomeindividuals. c)Spermandoocytesarehaploid d)escaperareembryostheyescapedfromexpressioneventhoughtheycontainedmaternal transgene.Theyusedpolymorphismanddevelopedprimerswhichboundtoexonsthatdidnot showdifferentialexpressionofitag6maternalandpaternalsituations.

9)Theylookedattrimethylationofh3k9anddidaChiPexperiementwithinvertedHOXD mutantsusingantibodiesdirectedandrepressivemarkers(H3K9)Thepurposewastosee whetherthesemarkersplayedaroleindownregulationoflacZ. 13)Regulatoryeffectsofimprinting: