摘要
为了探讨敲除生长分化因子11(gdf11)基因对小鼠生长性状的影响,本试验根据gdf 11基因序列设计向导RNA(sgRNA)并利用规律成蔟的短间隔回文重复序列和CRISPR相关蛋白(CRISPR/Cas9)基因编辑系统对该基因进行敲除,利用显微注射的方法获得gdf 11敲除型F0代小鼠,通过聚合酶链式反应(PCR)和测序鉴定子代小鼠基因型。繁育后观察F1代小鼠形态,并采用骨架染色方法观察不同基因型小鼠的性状。结果显示,成功构建了gdf 11基因敲除小鼠,并且通过繁育得到了野生型(gdf 11^(+/+))、杂合型(gdf 11^(+/-))和纯合型(gdf 11^(-/-))3种基因型的小鼠;形态观察发现,与野生型小鼠相比,纯合型小鼠在围产期死亡且尾巴是截断的,骨架染色显示,野生型小鼠有13个胸椎和6个腰椎,纯合型小鼠有18个胸椎和9个腰椎。结果表明,敲除gdf 11基因可能会影响小鼠胚胎和中轴骨发育从而改变小鼠的生长性状。
In order to explore the effect of knocking out the growth differentiation factor 11(gdf11)gene on the growth traits of mice,this study designed a small guide RNA(sgRNA),and knocked out the gene by using the regularly formed short palindrome repeats and CRISPR-associated proteins(CRISPR/Cas9)gene editing system.The F0 generation mice of gdf 11 knockout type were obtained by microinjection.The genotypes of the offspring mice were identified by polymerase chain reaction(PCR)and sequencing.After breeding,the F1 generation mice were observed and the traits of mice with different genotypes were observed by skeletal staining.The results showed that the gdf 11 gene knockout mice were successfully constructed,and the wild-type(gdf 11^(+/+)),heterozygous(gdf 11^(+/-))and homozygous(gdf 11^(-/-))mice were obtained through breeding.Compared with wild-type mice,homozygous mice died in perinatal period and their tails were truncated.Skeleton staining showed that wild-type mice had 13 thoracic vertebrae and 6 lumbar vertebrae,while homozygous mice had 18 thoracic vertebrae and 9 lumbar vertebrae.These results indicate that the knock-out of gdf 11 gene may affect the embryonic and axial bone development in mice,and thus change their growth traits.
作者
王悦
李村院
刘壮
安明轩
李晓悦
胡圣伟
倪伟
WANG Yue;LI Cun-yuan;LIU Zhuang;AN Ming-xuan;LI Xiao-yue;HU Sheng-wei;NI Wei(College of Life Sciences,Shihezi University,Shihezi 832003,China;College of Animal Science and Technology,Shihezi University,Shihezi 832003,China)
出处
《中国兽医杂志》
CAS
北大核心
2023年第4期7-12,共6页
Chinese Journal of Veterinary Medicine
基金
新疆生产建设兵团国际科技合作项目(2018BC011)
新疆生产建设兵团科技攻关计划项目(2019AB034,2021CB033)。
