User:Pakpoom Subsoontorn/Notebook/Genetically Encoded Memory/2008/10/28

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PhiC31 Host: Streptomyces lividans Action: tyrosine integrase Structure: length:: 613 aa crystal structure is unknown. Target sequence: attB:: has two inverted repeated sequence (core-type) flanking an overlap region (TTG). Minimal attB is 34 bps while minimal attP is 39 bp for e.coli intramolecular integration. attP:: has core-type sites and overlap region, without an arm-type site. --See the schematic sequence of attB and attP at (1-f1). See Groth-2000 for experiment for minimal sequence. --See Thyagarajan-2001 for integration of attB and attP above background/pseudointegration site ~attP in human genome (these could be very cool model experiment!) Change specificity: Mutation/directed evolution could change specificity to pseudo attP site (HpsA, 44% identity) Mechanism: Excision: Excisionase has not yet been found. mutant/hybrid directed evolution for targeting human's site Non-natural host: Integration works in E.coli, mammalian and in vitro, Mouse embryos Hollis et al 2003 Bovine fiboblase--Ma et al 2006 Drosophila--Groth et al 2004 Aedes aegyptus--Nimmo et al 2006 Silk worm--Nakayama et al 2006 cassette exchange: Drosphila --Bateman et al 2006, fission yeast--Thomson et al 2001, mouse embryonic stem cell--Belteki et al 2003 Xenopus laevis--Allen&Weeks 2005 chloroplast transformation--Lutz et al 2004 gene therapy example--table1 for ref-1 References 1Calos MP. The PhiC31 Integrase System for Gene Therapy. Current gene Therapy, 2006, 6, 633-645.