\u30d5\u30e9\u30f3\u30b9\u8996\u899a\u7814\u7a76\u6240\u30fb\u30bd\u30eb\u30dc\u30f3\u30cc\u5927\u5b66Livet\u7814\u7a76\u5ba4\u306e\u9688\u5143\u62d3\u99ac\u535a\u58eb\u3092\u304a\u62db\u304d\u3057\u3066\u30bb\u30df\u30ca\u30fc\u3092\u958b\u50ac\u81f4\u3057\u307e\u3059\u3002\u795e\u7d4c\u767a\u751f\u7814\u7a76\u306e\u305f\u3081\u306e\u795e\u7d4c\u7d30\u80de\u591a\u8272\u6a19\u8b58\u6cd5\u304a\u3088\u3073\u30a4\u30e1\u30fc\u30b8\u30f3\u30b0\u6280\u8853\u306b\u3064\u3044\u3066\u3054\u8b1b\u6f14\u3044\u305f\u3060\u304d\u307e\u3059\u3002\u3075\u308b\u3063\u3066\u3054\u53c2\u52a0\u4e0b\u3055\u3044\u3002\u5b66\u90e8\u5b66\u751f\u30fb\u5927\u5b66\u9662\u751f\u306e\u53c2\u52a0\u3082\u6b53\u8fce\u3067\u3059\u3002\u30bb\u30df\u30ca\u30fc\u306f\u82f1\u8a9e\u3067\u3059\u304c\u3001\u8cea\u7591\u306f\u65e5\u672c\u8a9e\u3067\u3082\u82f1\u8a9e\u3067\u3082\u53ef\u80fd\u3067\u3059\u3002<\/p>\n
\u6f14\u8005\uff1a\u3000\u9688\u5143\u3000\u62d3\u99ac\u3000\u535a\u58eb<\/p>\n
\u6240\u5c5e\uff1a\u3000Jean LIVET Lab, Institut de la Vision-Sorbonne Universit\u00e9, France<\/p>\n
\u30bf\u30a4\u30c8\u30eb\uff1a\u3000A novel genome integrating vector system for direct transgenesis
\nin neural development<\/p>\n
\u65e5\u6642\uff1a\u30002019\u5e7412\u67085\u65e5\uff08\u6728\uff0917:00\uff5e18:00
\n\u5834\u6240\uff1a\u3000\u533b\u5b66\u90e8\u57fa\u790e\u7814\u7a76A\u68df1\u968e\u3000\u7b2c2\u8b1b\u7fa9\u5ba4\uff08\u751f\u5354\u5411\u304b\u3044\uff09<\/p>\n
Abstract: In developmental neuroscience, direct transfection of neural stem cells in vivo with genome-integrating vectors is increasingly used as an alternative to the generation of transgenic animals. However, current genome-integrative vectors present important limitations: retroviral vectors traditionally used in cell lineage studies are cumbersome to produce and limited in cargo capacity. Naked DNA vectors are considerably simpler to use and can accept large transgenes, but they enable expression from non-integrated episomes in addition to that of integrated transgenes. This creates a transient burst of expression that prevents reliable identification of transgenic cells and can cause transgene leakiness and toxicity. We bypass this issue with a novel expression strategy termed \u201ciOn\u201d (integration-coupled On expression) switch, enabling to activate DNA transgenes introduced by standard transfection procedures as they integrate into the host genome, while episomal transgenes remain silent. Toxic and non-physiological effects due to high episomal expression are thus avoided. Marker expression solely reflects the activity of integrated transgenes, enabling straightforward identification and analysis of integration events directly after transfection.
\nWe demonstrate the efficiency of the iOn strategy for stable additive transgenesis in neural progenitors in the embryonic cortex, retina and spinal cord of mice and chicken. In these systems, we show that iOn vectors coupled with fluorescent color reporters make an ideal tool for cell lineage tracing and open the way to functional mosaic analysis through simple and rapid somatic transfection procedures. iOn vectors also drive leak-proof genome integration-dependent Cre-lox recombination, which we use to reveal the clonal output of a genetically identified subtype of retinal progenitors in the developing chick retina. We also show that iOn vectors provide an efficient means to stably transfect and rapidly isolate clones of stem\/pluripotent cells in vitro. These results establish iOn as a highly efficient and versatile strategy for \u201cdirect transgenesis\u201d in any model accessible to transfection.<\/p>\n
\u4e16\u8a71\u4eba\uff1a\u3000\u533b\u5b66\u7814\u7a76\u9662\u30fb\u75be\u60a3\u60c5\u5831\u7814\u7a76\u5206\u91ce\u3000\u4eca\u4e95\u3000\u731b
\n#\u7b2c13\u56de\u75be\u60a3\u60c5\u5831\u7814\u7a76\u5206\u91ce\u4e3b\u50ac\uff08\u7b2c17\u56de\u4eca\u4e95\u7814\u4e3b\u50ac\uff09\u516c\u958b\u30bb\u30df\u30ca\u30fc<\/p>\n","protected":false},"excerpt":{"rendered":"
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