Gene-targeting and Animal Model Unit

在过去的一年里，我们的核心帮助NHLBI研究人员成功地培育了十几个转基因和基因敲除小鼠品系。其中四个项目是使用最近开发的靶向转基因(也称为定点转基因)方法完成的。我们的单位还投入了相当大的努力来开发快速发展的基因组工程技术中的新能力。我们已经成功地使用了锌指核酸酶(ZFN)、TALEN和CRISPR/Cas9方法来编辑诱导多能干细胞(IPSC)系的基因组。我们还成功地使用ZFN方法制造了基因敲除小鼠系，目前正在使用TALEN和CRISPR/Cas9方法来产生基因敲除、条件基因敲除和敲除小鼠系。我们还参与了一系列与干细胞相关的项目，包括动物IPSC/ESC来源、畸胎瘤形成实验、干细胞移植的小鼠模型和组织工程。在过去的一年里，我们为NHLBI内外的科学家提供了大量的咨询和技术援助，帮助他们进行小鼠分子遗传学研究，包括设计DNA结构、搜索数据库、收集不同阶段的胚胎、获得小鼠胚胎成纤维细胞(MEF)、收集小鼠器官，以及使用冷冻精子重建小鼠系。我也是NHLBI动物护理和使用委员会(ACUC)的成员，负责审查动物规程和进行动物设施检查。

In the past year, our core has assisted NHLBI investigators in successfully generating more than one dozen transgenic and knockout mouse lines. Four of these projects were completed using the recently developed targeted transgenic (also called site-specific transgenic) method. Our unit has also devoted a significant portion of our effort on developing new capabilities in the rapidly advancing genome engineering technologies. We have successfully used the ZFN (zinc finger nuclease), TALEN, and CRISPR/Cas9, method to edit the genome of induced pluripotent stem cell (iPSC) lines. We have also succeeded in producing knockout mouse lines using the ZFN method, and are currently using the TALEN and CRISPR/Cas9 methods to generate knockout, conditional knockout, and knockin mouse lines. We have also been involved in a range of stem cell related projects, including animal iPSC/ESC derivation, teratoma formation assay, mouse models of stem cell transplantation, and tissue engineering. Throughout the year, we have provided numerous consultations and technical assistances to scientists with in and out of NHLBI to assist them in conducting mouse molecular genetic research, including designing DNA constructs, searching database, collecting various stages of embryos, deriving mouse embryonic fibroblasts (MEF), collecting mouse organs, and reconstituting mouse lines using cryopreserved sperm. I am also a member of the NHLBI Animal Care and Use Committee (ACUC) for reviewing animal protocols and conducting animal facility inspections.