Transgenic Core

在过去的一年里，我们的核心已经帮助NHLBI的研究人员成功地产生了十多个转基因和敲除小鼠系。在这一年中，我们的核心已经建立了一种新的能力，即通过将转基因与Phi31重组酶共同注射到受精卵的原核中，将转基因插入到特定的基因组位点。我们还成功地从早期小鼠胚胎中获得小鼠胚胎干细胞（ESC）和系统干细胞（EpiSC）系。我们的核心也投入了大量的精力来开发诱导多能干细胞（iPSC）相关的能力。我们已经在这一领域获得了广泛的技能，包括iPSC的衍生，通过胚体形成和畸胎瘤形成的随机分化，核型，以及向心肌细胞和神经元谱系的定向分化。我们已经协助几个实验室获得了许多小鼠iPSC系，并成功地从表达gfp的转基因猪中获得了iPSC系。我们还成功地将报告基因定位到来自NIH患者的人类iPSCs的安全港位点（AAVS1）。我们还将小鼠iPSCs和造血干细胞（hsc）微注射到从不能形成血统的敲除小鼠品系收集的囊胚中。在过去的一年中，我们为NHLBI内外的科学家提供了大量的咨询和技术援助，以协助他们进行小鼠分子遗传学研究，包括设计DNA结构，检索数据库，收集胚胎的各个阶段，获得小鼠胚胎成纤维细胞（MEF），收集小鼠器官，以及使用冷冻保存的精子重建小鼠系。我也是NHLBI动物护理和使用委员会（ACUC）的成员，负责审查动物协议并进行动物设施检查。

In the past year, our core has assisted NHLBI investigators in successfully generating more than one dozen transgenic and knockout mouse lines. One new capability our core has established during this year is the generation of targeted transgenic mouse lines, which is to insert a transgene into a specific genomic locus by co-injecting the transgene with the Phi31 recombinase into the pronuclei of fertilized eggs. We have also succeeded in deriving mouse embryonic stem cell (ESC) and epistem cell (EpiSC) lines from early mouse embryos. Our core has also devoted a substantial portion of our efforts on developing induced pluripotent stem cell (iPSC) related capabilities. We have gained a broad range of skills in this area, including iPSC derivation, random differentiation through embroid body formation and teratoma formation, karyotyping, and directed differentiation into cardiomyocyte and neuronal lineage. We have assisted several laboratories in deriving a number of mouse iPSC lines, and succeeded in deriving iPSC lines from a GFP-expressing transgenic pig. We have also succeeded in targeting reporter genes into a safe harbor locus (AAVS1) of human iPSCs derived from NIH patients. We aslo microinjected mouse iPSCs and hematopoietic stem cells (HSCs) into blastocysts collected from a knockout mouse strain, which is incapable of forming blood lineage. 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.