Abstract:
:Creating transgenic mice is an important technology for genetic studies and is currently performed by pronuclear microinjection of plasmid DNA into fertilized eggs. Since survival of injected embryos and integration of plasmid DNA are not efficient, total efficiency is only around 3% with a standard protocol. To circumvent this problem, here we describe a novel transgenesis method, the Tol2-mediated cytoplasmic injection method (Tol2:CI). We injected a foreign DNA cloned in a Tol2-transposon vector together with the transposase mRNA into the cytoplasm of fertilized eggs. As expected, the survival rate of the injected embryos was increased drastically. Also, the foreign DNA was transposed from the plasmid to the genome and transmitted to the next generation very efficiently. Together, the overall transgenic efficiency became more than 20%. Considering its simplicity and perfect compatibility with existing pronuclear microinjection facilities, we propose that the Tol2:CI method is applicable to high throughput functional genomics studies.
journal_name
Genomicsjournal_title
Genomicsauthors
Sumiyama K,Kawakami K,Yagita Kdoi
10.1016/j.ygeno.2010.02.006subject
Has Abstractpub_date
2010-05-01 00:00:00pages
306-11issue
5eissn
0888-7543issn
1089-8646pii
S0888-7543(10)00039-Xjournal_volume
95pub_type
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