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Eriko Toyoda Aya Kurosawa Noritaka Adachi 《Biochemical and biophysical research communications》2009,382(3):492-496
DNA transfection can be too inefficient to establish a desired number of stable transfectants, particularly in lymphocytes; however, this could be circumvented by increasing the absolute frequency of random integration. In this paper, we show that treating cells with topoisomerase II inhibitor following electroporation greatly (∼10- to 20-fold) enhances random integration of input DNA in a human pre-B lymphocyte cell line, Nalm-6. With the use of various kinds of topoisomerase II-targeting agents, we also present evidence that topoisomerase IIα inhibition is critical for the enhancement of random integration, while the contribution of topoisomerase IIβ may be negligible. As topoisomerase IIα is highly expressed in vigorously growing cells, our results show that topoisomerase IIα inhibition provides a promising way of enhancing random integration in virtually all cultured cell lines. 相似文献
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Paola Sebastiani Monty Montano Annibale Puca Nadia Solovieff Toshio Kojima Meng C. Wang Efthymia Melista Micah Meltzer Sylvia E. J. Fischer Stacy Andersen Stephen H. Hartley Amanda Sedgewick Yasumichi Arai Aviv Bergman Nir Barzilai Dellara F. Terry Alberto Riva Chiara Viviani Anselmi Alberto Malovini Aya Kitamoto Motoji Sawabe Tomio Arai Yasuyuki Gondo Martin H. Steinberg Nobuyoshi Hirose Gil Atzmon Gary Ruvkun Clinton T. Baldwin Thomas T. Perls 《PloS one》2009,4(12)
Background
The strong familiality of living to extreme ages suggests that human longevity is genetically regulated. The majority of genes found thus far to be associated with longevity primarily function in lipoprotein metabolism and insulin/IGF-1 signaling. There are likely many more genetic modifiers of human longevity that remain to be discovered.Methodology/Principal Findings
Here, we first show that 18 single nucleotide polymorphisms (SNPs) in the RNA editing genes ADARB1 and ADARB2 are associated with extreme old age in a U.S. based study of centenarians, the New England Centenarian Study. We describe replications of these findings in three independently conducted centenarian studies with different genetic backgrounds (Italian, Ashkenazi Jewish and Japanese) that collectively support an association of ADARB1 and ADARB2 with longevity. Some SNPs in ADARB2 replicate consistently in the four populations and suggest a strong effect that is independent of the different genetic backgrounds and environments. To evaluate the functional association of these genes with lifespan, we demonstrate that inactivation of their orthologues adr-1 and adr-2 in C. elegans reduces median survival by 50%. We further demonstrate that inactivation of the argonaute gene, rde-1, a critical regulator of RNA interference, completely restores lifespan to normal levels in the context of adr-1 and adr-2 loss of function.Conclusions/Significance
Our results suggest that RNA editors may be an important regulator of aging in humans and that, when evaluated in C. elegans, this pathway may interact with the RNA interference machinery to regulate lifespan. 相似文献296.
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Mikihiro Yunoki Hiroyuki Tanaka Takeru Urayama Yuta Kanai Aya Nishida Maki Yoshikawa Yuji Ohkubo Yoshiyasu Kawabata Katsuro Hagiwara Kazuyoshi Ikuta 《Biologicals》2010,38(2):311-313
The evaluation of the removal efficacy during manufacturing is important for the risk assessment of plasma products with respect to possible contamination by infectious prions, as recently reported in several papers on the potential for prion transmission through plasma products. Here, we evaluated a virus removal filter which has 15 nm pores. An antithrombin sample immediately prior to nano-filtration was spiked with prion material prepared in two different ways. The removal (log reduction factor) of prion infectivity using animal bioassays was ≥4.72 and 4.00 in two independent filtrations. However, infectivity was detected in both the pellet and supernatant following ultracentrifugation of the 15 nm filtered samples, indicating difficulty in complete removal. The data supports the conclusion that a certain amount of infectious prion protein is present as a smaller and/or soluble form (less than ~15 nm in diameter). 相似文献
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Akito Imai Mai Ohtani Asami Nara Anna Tsukakoshi Aya Narita Hideki Hirakawa Shusei Sato Norio Suganuma 《Physiologia plantarum》2020,168(3):590-600
Nucleoporins are components of the nuclear pore complexes, channels that regulate the transport of macromolecules between the nucleus and cytoplasm. The nucleoporin GLE1 (GLFG lethal1) functions in the export of messenger RNAs containing poly(A) tails from the nucleus into the cytoplasm. Here we investigated a mutant of the model legume Lotus japonicus that was defective in GLE1, which we designated Ljgle1. The growth of Ljgle1 was retarded under symbiotic association with rhizobia, and the nitrogen-fixation activities of the nodules were around one-third of those in the wild-type plant. The growth of Ljgle1 was not substantialy recovered by supplemention of combined nitrogen. Nodules formed on the Ljgle1 were smaller than those on the wild-type and colored faint pink. The numbers of infected cells of nodules on the Ljgle1 were smaller than on the wild-type plant, and the former cells remained undeveloped. Rhizobia in the cells of the Ljgle1 exhibited disordered forms, and the symbiosome membrane was closely attached to the bacterial membrane. These results indicate that GLE1 plays a distinct role in the symbiotic association between legumes and rhizobia. 相似文献
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