Balkow Aileen, Hoffmann Linda S, Klepac Katarina, Glöde Anja, Gnad Thorsten, Zimmermann Katrin, Pfeifer Alexander
Institute of Pharmacology and Toxicology, University Hospital Bonn, University of Bonn, 53127 Bonn, Germany.
Research Training Group 1873, University of Bonn, 53127 Bonn, Germany.
J Biol Methods. 2016 Jul 16;3(3):e48. doi: 10.14440/jbm.2016.123. eCollection 2016.
Brown adipose tissue is a special type of fat contributing to energy expenditure in human newborns and adults. Moreover, subcutaneous white adipose tissue has a high capacity to adapt an energy-consuming, brown-like/beige phenotype. Here, we developed an easy to handle and fast to accomplish method to efficiently transfer genes into brown and beige fat pads . Lentiviral vectors are directly injected into the target fat pad of anesthetized mice through a small incision using a modified, small needle connected to a microsyringe, which is well suited for infiltration of adipose tissues. Expression of the target gene can be detected in brown/beige fat one week after injection. The method can be applied within minutes to efficiently deliver transgenes into subcutaneous adipose tissues. Thus, this protocol allows for studying genes of interest in a timely manner in murine brown/beige fat and could potentially lead to new gene therapies for obesity.
棕色脂肪组织是一种特殊类型的脂肪，对人类新生儿和成年人的能量消耗有贡献。此外，皮下白色脂肪组织具有很高的能力来适应耗能的、类似棕色/米色的表型。在此，我们开发了一种易于操作且快速完成的方法，可有效地将基因导入棕色和米色脂肪垫。通过使用连接到微量注射器的改良小针，经小切口将慢病毒载体直接注射到麻醉小鼠的目标脂肪垫中，这种方法非常适合脂肪组织的浸润。注射一周后可在棕色/米色脂肪中检测到目标基因的表达。该方法可在数分钟内应用，以有效地将转基因递送至皮下脂肪组织。因此，该方案能够及时在小鼠棕色/米色脂肪中研究感兴趣的基因，并有可能带来针对肥胖症的新基因疗法。
