Genome-wide target profiling of piggyBac and Tol2 in HEK 293: pros and cons for gene discovery and gene therapy

Hdl Handle:
http://hdl.handle.net/10675.2/736
Title:
Genome-wide target profiling of piggyBac and Tol2 in HEK 293: pros and cons for gene discovery and gene therapy
Authors:
Meir, Yaa-Jyuhn J; Weirauch, Matthew T; Yang, Herng-Shing; Chung, Pei-Cheng; Yu, Robert K.; Wu, Sareina C-Y
Abstract:
Background: DNA transposons have emerged as indispensible tools for manipulating vertebrate genomes with applications ranging from insertional mutagenesis and transgenesis to gene therapy. To fully explore the potential of two highly active DNA transposons, piggyBac and Tol2, as mammalian genetic tools, we have conducted a side-by-side comparison of the two transposon systems in the same setting to evaluate their advantages and disadvantages for use in gene therapy and gene discovery.; Results: We have observed that (1) the Tol2 transposase (but not piggyBac) is highly sensitive to molecular engineering; (2) the piggyBac donor with only the 40 bp 3'-and 67 bp 5'-terminal repeat domain is sufficient for effective transposition; and (3) a small amount of piggyBac transposases results in robust transposition suggesting the piggyBac transpospase is highly active. Performing genome-wide target profiling on data sets obtained by retrieving chromosomal targeting sequences from individual clones, we have identified several piggyBac and Tol2 hotspots and observed that (4) piggyBac and Tol2 display a clear difference in targeting preferences in the human genome. Finally, we have observed that (5) only sites with a particular sequence context can be targeted by either piggyBac or Tol2.; Conclusions: The non-overlapping targeting preference of piggyBac and Tol2 makes them complementary research tools for manipulating mammalian genomes. PiggyBac is the most promising transposon-based vector system for achieving site-specific targeting of therapeutic genes due to the flexibility of its transposase for being molecularly engineered. Insights from this study will provide a basis for engineering piggyBac transposases to achieve site-specific therapeutic gene targeting.
Citation:
BMC Biotechnol. 2011 Mar 30; 11:28
Issue Date:
30-Mar-2011
URI:
http://hdl.handle.net/10675.2/736
DOI:
10.1186/1472-6750-11-28
PubMed ID:
21447194
PubMed Central ID:
PMC3078864
Type:
Article
ISSN:
1472-6750
Appears in Collections:
Institute of Molecular Medicine and Genetics: Faculty Research and Presentations

Full metadata record

DC FieldValue Language
dc.contributor.authorMeir, Yaa-Jyuhn Jen_US
dc.contributor.authorWeirauch, Matthew Ten_US
dc.contributor.authorYang, Herng-Shingen_US
dc.contributor.authorChung, Pei-Chengen_US
dc.contributor.authorYu, Robert K.en_US
dc.contributor.authorWu, Sareina C-Yen_US
dc.date.accessioned2012-10-26T20:27:52Z-
dc.date.available2012-10-26T20:27:52Z-
dc.date.issued2011-03-30en_US
dc.identifier.citationBMC Biotechnol. 2011 Mar 30; 11:28en_US
dc.identifier.issn1472-6750en_US
dc.identifier.pmid21447194en_US
dc.identifier.doi10.1186/1472-6750-11-28en_US
dc.identifier.urihttp://hdl.handle.net/10675.2/736-
dc.description.abstractBackground: DNA transposons have emerged as indispensible tools for manipulating vertebrate genomes with applications ranging from insertional mutagenesis and transgenesis to gene therapy. To fully explore the potential of two highly active DNA transposons, piggyBac and Tol2, as mammalian genetic tools, we have conducted a side-by-side comparison of the two transposon systems in the same setting to evaluate their advantages and disadvantages for use in gene therapy and gene discovery.en_US
dc.description.abstractResults: We have observed that (1) the Tol2 transposase (but not piggyBac) is highly sensitive to molecular engineering; (2) the piggyBac donor with only the 40 bp 3'-and 67 bp 5'-terminal repeat domain is sufficient for effective transposition; and (3) a small amount of piggyBac transposases results in robust transposition suggesting the piggyBac transpospase is highly active. Performing genome-wide target profiling on data sets obtained by retrieving chromosomal targeting sequences from individual clones, we have identified several piggyBac and Tol2 hotspots and observed that (4) piggyBac and Tol2 display a clear difference in targeting preferences in the human genome. Finally, we have observed that (5) only sites with a particular sequence context can be targeted by either piggyBac or Tol2.en_US
dc.description.abstractConclusions: The non-overlapping targeting preference of piggyBac and Tol2 makes them complementary research tools for manipulating mammalian genomes. PiggyBac is the most promising transposon-based vector system for achieving site-specific targeting of therapeutic genes due to the flexibility of its transposase for being molecularly engineered. Insights from this study will provide a basis for engineering piggyBac transposases to achieve site-specific therapeutic gene targeting.en_US
dc.rightsCopyright ©2011 Meir et al; licensee BioMed Central Ltd.en_US
dc.subjectResearch Articleen_US
dc.titleGenome-wide target profiling of piggyBac and Tol2 in HEK 293: pros and cons for gene discovery and gene therapyen_US
dc.typeArticleen_US
dc.identifier.pmcidPMC3078864en_US
dc.contributor.corporatenameInstitute of Molecular Medicine and Genetics-
dc.contributor.corporatenameInstitute of Neuroscience-

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