摘要
目的:重组腺相关病毒载体(rAAV)是治疗遗传性疾病和慢性病最有前途的基因载体,本文研究重组腺相关病毒介导的基因转染的遗传效应。方法:将成年的2月龄雄性昆明小鼠分为三组,分别自尾静脉注入rAAV-D(+)-HK、rAAV-D(+)-GFP病毒颗粒和生理盐水,两周后将其以1:1的比例与成年的雌性昆明小鼠合笼。观察各组小鼠在一周内的受孕率、各组产仔情况、第一代及其子代的主要脏器组织形态学特性、HK基因在第一代及其子代的表达情况,以及HK基因的整合情况等。结果:重组腺相关病毒对基因转染小鼠的受孕率、产仔情况无影响;重组腺相关病毒携带的目的基因可以在第一代和第二代小鼠中实现稳定表达,并可整合到第一代和第二代小鼠的基因组中,基因转染的第一代和第二代小鼠的主要脏器病理学检查均未见明显异常;重组腺相关病毒并未影响第二代小鼠的生长发育。结论:重组腺相关病毒载体可通过昆明小鼠的血-睾屏障,在第二代小鼠中实现稳定表达。且在第一代和第二代昆明小鼠中均未观测到明显的毒性反应。
Objective Recombinant adeno-associated virus is the most promising gene vector that is used to treat genetic and chronic diseases. The present study investigated the genetic effects of gene delivery mediated by recombinant adeno-associated virus （rAAV）. Method Adult male Kunming （KM）mice were divided into three groups and injected intravenously in the tail vein with rAAV-D （ ＋ ）-HK, rAAV-D （ ＋ ）-GFP viral particles and saline respectively. Two weeks later, we matched the male mice with adult female mice in the ratio of 1 ： 1. Then we observed the rate of pregnancy in a week and production. We studied pathological changes of the major organs of the first and second generation and the expression and integration of HK gene. Results rAAV did not affect the pregnancy and production of the treated mice. Target gene carried by rAAV can express stably in the first and second generation of KM mice, and integrate into the genomes of the first and second generation. No significant abnormalities were found in the pathological study of the major organs in the first and second generation of the mice. rAAV did not affect the growth and development of the second generation of the mice. Conclusion rAAV can pass the blood-testis barrier of KM mice and mediate stable expression of target gene in second generation of KM mice. No significant toxic actions are found in the first and second generation of KM mice.
出处
《中国分子心脏病学杂志》
CAS
2003年第2期72-78,共7页
Molecular Cardiology of China
基金
国家高技术研究发展计划(863计划
No.2001AA217121)
武汉市科委资助项目(No.997005135G)
