Mouse Genomics Resource Laboratory, National Institute of Genetics, Mishima, 411-8540, Japan.
Laboratory for Mouse Genetic Engineering, RIKEN Center for Biosystems Dynamics Research, Osaka, 565-0871, Japan.
Sci Rep. 2022 Aug 15;12(1):13821. doi: 10.1038/s41598-022-17776-x.
Wild mouse strains have been used for many research studies, because of the high level of inter-strain genetic and phenotypic variations in them, in addition to the characteristic phenotype maintained from wild mice. However, since application of the current genetic engineering method on wild strains is not easy, there are limited studies that have attempted to apply gene modification techniques in wild strains. Recently, i-GONAD, a new method for genome editing that does not involve any ex vivo manipulation of unfertilized or fertilized eggs has been reported. We applied i-GONAD method for genome editing on a series of wild strains and showed that genome editing is efficiently possible using this method. We successfully made genetically engineered mice in seven out of the nine wild strains. Moreover, we believe that it is still possible to apply milder conditions and improve the efficiencies for the remaining two strains. These results will open avenues for studying the genetic basis of various phenotypes that are characteristic to wild strains. Furthermore, applying i-GONAD will be also useful for other mouse resources in which genetic manipulation is difficult using the method of microinjection into fertilized eggs.
野生鼠种已被广泛应用于多项研究中，这是因为它们具有高水平的种间遗传和表型变异，此外还保留了来自野生鼠的特征表型。然而，由于当前的遗传工程方法在野生品系上的应用并不容易，因此只有有限的研究尝试将基因修饰技术应用于野生品系。最近，一种新的基因组编辑方法 i-GONAD 被报道，该方法不涉及对未受精或受精卵的任何体外操作。我们应用 i-GONAD 方法对一系列野生品系进行了基因组编辑，结果表明该方法可以有效地进行基因组编辑。我们在其中的 9 个野生品系中的 7 个中成功地制造了基因工程小鼠。此外，我们认为对于其余的两个品系，仍然有可能应用更温和的条件并提高效率。这些结果将为研究野生品系特有各种表型的遗传基础开辟途径。此外，应用 i-GONAD 对于其他使用受精卵微注射方法进行遗传操作困难的小鼠资源也将是有用的。
