Schlesinger-Laufer Michal, Douvdevany Guy, Haimovich-Caspi Lilac, Zohar Yaniv, Shofty Rona, Kehat Izhak
The Pre-Clinical Research Authority Unit, The Technion, Israel Institute of Technology, 1 Efron Street, P.O. Box 9697, Haifa, 3109601, Israel.
Faculty of Medicine, Technion - Israel Institute of Technology, 1 Efron Street, P.O. Box 9697, Haifa, 3109601, Israel.
F1000Res. 2020 Dec 10;9. doi: 10.12688/f1000research.27675.1. eCollection 2020.
Heart failure is a major health problem and progress in this field relies on better understanding of the mechanisms and development of novel therapeutics using animal models. The rat may be preferable to the mouse as a cardiovascular disease model due to its closer physiology to humans and due to its large size that facilitates surgical and monitoring procedures. However, unlike the mouse, genetic manipulation of the rat genome is challenging. Here we developed a simple, refined, and robust cardiac-specific rat transgenic model based on an adeno-associated virus (AAV) 9 containing a cardiac troponin T promoter. This model uses a single intraperitoneal injection of AAV and does not require special expertise or equipment. : We characterize the AAV dose required to achieve a high cardiac specific level of expression of a transgene in the rat heart using a single intraperitoneal injection to neonates. We show that at this AAV dose GFP expression does not result in hypertrophy, a change in cardiac function or other evidence for toxicity. The model shown here allows easy and fast transgenic based disease modeling of cardiovascular disease in the rat heart, and can also potentially be expanded to deliver Cas9 and gRNAs or to deliver small hairpin (sh)RNAs to also achieve gene knockouts and knockdown in the rat heart.
心力衰竭是一个重大的健康问题，该领域的进展依赖于对发病机制的更好理解以及利用动物模型开发新型治疗方法。大鼠作为心血管疾病模型可能比小鼠更具优势，因为其生理机能与人类更为接近，且体型较大便于进行手术和监测操作。然而，与小鼠不同的是，对大鼠基因组进行基因操作具有挑战性。在此，我们基于携带心肌肌钙蛋白T启动子的腺相关病毒（AAV）9开发了一种简单、精细且稳定的心脏特异性大鼠转基因模型。该模型通过单次腹腔注射AAV，无需特殊专业知识或设备。我们通过对新生大鼠进行单次腹腔注射，确定了在大鼠心脏中实现转基因高心脏特异性表达所需的AAV剂量。我们发现，在此AAV剂量下，绿色荧光蛋白（GFP）表达不会导致心肌肥大、心脏功能改变或其他毒性证据。本文展示的模型能够在大鼠心脏中轻松快速地基于转基因进行心血管疾病建模，并且还可能扩展用于递送Cas9和引导RNA（gRNA），或递送小发夹RNA（shRNA），以在大鼠心脏中实现基因敲除和基因敲低。
