Mutant Mos1 mariner transposons are hyperactive in Aedes aegypti

Pledger, David; Coates, Craig J.

doi:10.1016/j.ibmb.2005.06.002

Cited by 22 publications

(20 citation statements)

References 54 publications

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“…There is currently work being done with PB transposase to increase the transpositional efficiencies to that of retroviral vectors. There are methods to accomplish this goal such as PCR random mutagenesis or alanine substitutions utilizing mutagenic PCR primers (Goryshin and Reznikoff 1998;Yant et al 2004;Pledger and Coates 2005;Keravala et al 2006). One of us, Thomas Ryan, is pursuing PB active transgenesis in embryonic stem cells for the production of transgenic animals.…”

Section: Mechanisms To Improve Specificity and Efficiency Of Transfecmentioning

confidence: 99%

Active integration: new strategies for transgenesis

Shinohara

Kaminski²,

Segal

et al. 2007

Transgenic Res

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This paper presents novel methods for producing transgenic animals, with a further emphasis on how these techniques may someday be applied in gene therapy. There are several passive methods for transgenesis, such as pronuclear microinjection (PNI) and Intracytoplasmic Sperm Injection-Mediated Transgenesis (ICSI-Tr), which rely on the repair mechanisms of the host for transgene (tg) insertion. ICSI-Tr has been shown to be an effective means of creating transgenic animals with a transfection efficiency of approximately 45% of animals born. Furthermore, because this involves the injection of the transgene into the cytoplasm of oocytes during fertilization, limited mosaicism has traditionally occurred using this technique. Current active transgenesis techniques involve the use of viruses, such as disarmed retroviruses which can insert genes into the host genome. However, these methods are limited by the size of the sequence that can be inserted, high embryo mortality, and randomness of insertion. A novel active method has been developed which combines ICSI-Tr with recombinases or transposases to increase transfection efficiency. This technique has been termed "Active Transgenesis" to imply that the tg is inserted into the host genome by enzymes supplied into the oocyte during tg introduction. DNA based methods alleviate many of the costs and time associated with purifying enzyme. Further studies have shown that RNA can be used for the transposase source. Using RNA may prevent problems with continued transposase activity that can occur if a DNA transposase is integrated into the host genome. At present piggyBac is the most effective transposon for stable integration in mammalian systems and as further studies are done to elucidate modifications which improve piggyBac's specificity and efficacy, efficiency in creating transgenic animals should improve further. Subsequently, these methods may someday be used for gene therapy in humans.

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Section: Mechanisms To Improve Specificity and Efficiency Of Transfecmentioning

confidence: 99%

Active integration: new strategies for transgenesis

Shinohara

Kaminski²,

Segal

et al. 2007

Transgenic Res

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“…Their movement around the host genome has played a crucial role in shaping current genomes. Owing to their natural property of mobilizing around genomes via a ''cut-and-paste'' mechanism, DNA transposons have been routinely used as tools for genetic manipulation in lower organisms (3)(4)(5)(6). Only recently have scientists realized the potential value of transposons in mammalian transgenesis because most naturally occurring transposons are inactive and possibly were overlooked until recently because of the success of integrating viral vectors.…”

mentioning

confidence: 99%

piggyBac is a flexible and highly active transposon as compared to Sleeping Beauty , Tol2 , and Mos1 in mammalian cells

Meir

Coates

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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A nonviral vector for highly efficient site-specific integration would be desirable for many applications in transgenesis, including gene therapy. In this study we directly compared the genomic integration efficiencies of piggyBac, hyperactive Sleeping Beauty (SB11), Tol2, and Mos1 in four mammalian cell lines. piggyBac demonstrated significantly higher transposition activity in all cell lines whereas Mos1 had no activity. Furthermore, piggyBac transposase coupled to the GAL4 DNA-binding domain retains transposition activity whereas similarly manipulated gene products of Tol2 and SB11 were inactive. The high transposition activity of piggyBac and the flexibility for molecular modification of its transposase suggest the possibility of using it routinely for mammalian transgenesis.gene therapy ͉ site-specific ͉ transposase ͉ cancer ͉ mutagenesis

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“…Wild-type transposases are often suboptimal for transposition (Kunze et al, 1995;Lampe et al, 1999;Weil and Kunze, 2000;Zayed et al, 2004;Pledger and Coates, 2005;Kim and Morrison, 2009;Yusa et al, 2011;Lazarow et al, 2012). For example, mutation of three amino acids in the transposase of Himar1 led to a 4-to 50-fold increase in transposition activity (Lampe et al, 1999).…”

Section: The Low Copy Number Of Os3378 In Rice and Its Relatives Is Lmentioning

confidence: 99%

Transposition of a Rice Mutator-Like Element in the Yeast Saccharomyces cerevisiae

2015

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(N.J.)Mutator-like transposable elements (MULEs) are widespread in plants and are well known for their high transposition activity as well as their ability to duplicate and amplify host gene fragments. Despite their abundance and importance, few active MULEs have been identified. In this study, we demonstrated that a rice (Oryza sativa) MULE, Os3378, is capable of excising and reinserting in yeast (Saccharomyces cerevisiae), suggesting that yeast harbors all the host factors for the transposition of MULEs. The transposition activity induced by the wild-type transposase is low but can be altered by modification of the transposase sequence, including deletion, fusion, and substitution. Particularly, fusion of a fluorescent protein to the transposase enhanced the transposition activity, representing another approach to manipulate transposases. Moreover, we identified a critical region in the transposase where the net charge of the amino acids seems to be important for activity. Finally, transposition efficiency is also influenced by the element and its flanking sequences (i.e., small elements are more competent than their large counterparts). Perfect target site duplication is favorable, but not required, for precise excision. In addition to the potential application in functional genomics, this study provides the foundation for further studies of the transposition mechanism of MULEs.

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Mutant Mos1 mariner transposons are hyperactive in Aedes aegypti

Cited by 22 publications

References 54 publications

Active integration: new strategies for transgenesis

Active integration: new strategies for transgenesis

piggyBac is a flexible and highly active transposon as compared to Sleeping Beauty , Tol2 , and Mos1 in mammalian cells

Transposition of a Rice Mutator-Like Element in the Yeast Saccharomyces cerevisiae

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