经微注射质粒 DNA 构建体到雄原生殖细胞对粪类圆线虫进行遗传转化。
Heritable genetic transformation of Strongyloides stercoralis by microinjection of plasmid DNA constructs into the male germline.
机构信息
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, 3800 Spruce Street, Philadelphia, PA 19104, USA.
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, 3800 Spruce Street, Philadelphia, PA 19104, USA.
出版信息
Int J Parasitol. 2017 Aug;47(9):511-515. doi: 10.1016/j.ijpara.2017.04.003. Epub 2017 Jun 1.
Abstract
Heretofore, transgenesis in the parasitic nematode genus Strongyloides has relied on microinjecting transgene constructs into gonadal syncytia of free-living females. We now report transgenesis in Strongyloides stercoralis by microinjecting constructs into the syncytial testes of free-living males. Crosses of individual males microinjected with a construct encoding GFP with cohorts of 12 non-injected females produced a mean of 7.28±2.09 transgenic progeny. Progeny of males and females microinjected with distinct reporter constructs comprised 2.6%±0.7% of individuals expressing both paternal and maternal transgenes. Implications of this finding for deployment of CRISPR/Cas9 mutagenesis in Strongyloides spp. are discussed.
摘要
迄今为止，寄生线虫属 Strongyloides 的转基因依赖于将转基因构建体微注射到自由生活的雌性生殖合胞体中。我们现在报告了通过将构建体微注射到自由生活的雄性合胞体睾丸中转基因在 Strongyloides stercoralis 中的情况。用编码 GFP 的构建体单独微注射的个体雄性与 12 个未注射的雌性进行杂交，平均产生 7.28±2.09 个转基因后代。用不同报告基因构建体微注射的雄性和雌性后代中，同时表达父本和母本转基因的个体占 2.6%±0.7%。讨论了这一发现对 Strongyloides spp. 中 CRISPR/Cas9 诱变技术应用的影响。
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