Frøkjær-Jensen Christian
Howard Hughes Medical Institute, University of Utah, 257 S 1400 E RM 201, Salt Lake City, UT, 84112, USA.
Department of Biology, University of Utah, 257 S 1400 E RM 201, Salt Lake City, UT, 84112, USA.
Methods Mol Biol. 2015;1327:49-58. doi: 10.1007/978-1-4939-2842-2_5.
Transgenesis in model organisms is necessary to determine the function, expression, and subcellular localization of gene products. In Caenorhabditis elegans, injected DNA can be propagated as multicopy extrachromosomal arrays but transgenes in arrays are mosaic, over-expressed in some tissues and silenced in the germline. Here, a method to insert a transgene into a specific genomic location called Mos1-mediated single-copy insertion (MosSCI) is described. Single-copy insertion allows transgene expression at levels that approximate endogenous gene expression as well as expression in the germline.
在模式生物中进行转基因对于确定基因产物的功能、表达和亚细胞定位是必要的。在秀丽隐杆线虫中,注射的DNA可以作为多拷贝的染色体外阵列进行繁殖,但阵列中的转基因是嵌合的,在某些组织中过度表达,而在生殖系中沉默。在此,描述了一种将转基因插入特定基因组位置的方法,称为Mos1介导的单拷贝插入(MosSCI)。单拷贝插入使转基因能够以接近内源基因表达的水平表达,并且能够在生殖系中表达。