Phic31 integrase
WebApr 1, 2010 · For systems using the φC31-integrase, the recognition sites on the PP are attB and attP sites (the bacterial and phage attachment sites of the φC31-integrase with approximately 40 bp each). WebOct 28, 2024 · In one exemplary embodiment, the method disclosed herein selects a variant integrase (e.g., a phiC31 integrase) that can recognize a mutant attP sequence (e.g., SEQ ID NO: 3). The target mutant attP sequence is different from the wildtype attP site in more than 50% of the nucleotide residues, e.g., 26 out of 48 nucleotide residues are different.
Phic31 integrase
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WebJan 1, 2007 · The phiC31 integrase system represents a novel technology that opens up new possibilities for gene therapy. The phiC31 integrase can integrate introduced plasmid … WebJun 19, 2014 · The ΦC31 system uses the properties of the site-specific integrase from the bacteriophage ΦC31, which mediates recombination between an attachment site ( attB) that is normally present in the...
WebNational Center for Biotechnology Information WebMay 3, 2007 · ϕC31 integrase and several of its relatives are being widely used for precise engineering of complex genomes ( 1–8) and are emerging as promising new tools for gene therapy ( 9–15 ). In addition to being highly portable ϕC31 integrase is, unlike other recombinases used for genome manipulation such as Cre and Flp, unidirectional ( 7, 16, …
WebPhiC31 integrase can integrate a plasmid of any size, as a single copy, and requires no cofactors. The integrated transgenes are stably expressed and heritable. SBI’s PhiC31 … WebINTRODUCTION: The phiC31 integrase system is a phage-derived system that offers the ability to integrate plasmid DNA into the chromosomes at a subset of endogenous preferred locations associated with robust gene expression. Recent progress highlights the unique advantages of this system for in vivo gene therapy and for use in stem cells.
WebThe use of phiC31 integrase requires a defined sequence for insertion and is compatible with insertion of a single copy of the transgene. We describe the protocol we use to facilitate phiC31 integrase transgene insertion including the use of insulator sequences to reduce position effect disruption of transgene activity.
WebThe phiC31 integrase system has, for the first time, enabled transformation of flies with BAC-sized constructs . The integrase can be expressed from mRNA co-injected with the construct or from the genome under a germ-line specific promoter, the latter method being more efficient . Several dozens of ... assalierWebAug 11, 2011 · A generic PhiC31 integrase mediated exchange vector, pCB92, was assembled by modifying pExchange4-CB9 (derived from pRMCE [9]) with the insertion of a murine H19 insulator [20], obtained from pWHERE (Invivogen), together with a synthesized polylinker of unique sites to facilitate cloning. assali bpwWebThe present invention provides a method for producing single stranded DNA polynucleotides. In particular, the invention provides a method that utilises a DNA minicircle obtained from a parental minicircle plasmid as a template in an enzyme-mediated rolling circle amplification (RCA) reaction to generate a product that can be cleaved to provide … assa lietuvaWebNov 1, 2013 · Because the phiC31 integrase gene coding sequence contains one NcoI restriction site, the purified fragments were first cloned into the plasmid pMD19-T simple (Takara, Dalian, China) to generate plasmid T-Int, then the 1.9-kb target DNA fragment was recovered by incomplete digestion with NcoI and XhoI from T-Int, and inserted into the … assalightWebFeb 4, 2024 · We generated an integrated phiC31 integrase expressing strain flanked by attP sites that serves as a landing pad for integration of transgenes by recombination … assali autoWebJul 25, 2024 · A master cell line, with the landing pad inserted correctly in the H11 locus, was established. This master cell line was used for site-specific, irreversible recombination, catalyzed by PhiC31 integrase. Using this system, an integration efficiency of 97.7% was achieved with green fluorescent protein (GFP) after selection. lali y tini mueveteWebpUPD2 PhiC31 integrase (GB1561) was a gift from Diego Orzaez (Addgene plasmid # 160586 ; http://n2t.net/addgene:160586 ; RRID:Addgene_160586) For your References … assali el mehdi