RNAi assay in primary cells: a new method for gene function analysis in marine bivalve

doi:10.1007/s11033-012-1668-y

IOCAS-IR > 实验海洋生物学重点实验室

	RNAi assay in primary cells: a new method for gene function analysis in marine bivalve
	You, Yanan 1,2; Huan, Pin 1; Liu, Baozhong1 ; Liu, BZ (reprint author), Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China.
	2012-08-01
发表期刊	MOLECULAR BIOLOGY REPORTS
ISSN	0301-4851
卷号	39 期号:8 页码:8209-8216
文章类型	Article
摘要	RNA interference (RNAi) is an effective approach for gene function analysis, which is well developed in mammal cell lines. However, RNAi has rarely been reported in marine bivalve species. To provide support on functional analysis of bivalve genes, for the first time to our knowledge, we conducted RNAi assay on primary cell of clam Meretrix meretrix in this study. Firstly we explored the method of culturing primary cells of M. meretrix to ensure the cells to live at high activity for at least 2 weeks. Ferritin gene was chosen as the target gene and RNAi assay was conducted through soaking the primary cells of M. meretrix digestive gland in medium containing dsRNA of ferritin gene. Realtime PCR, western blot and immunocytochemistry analysis were used to analyze the inhibition of gene expression after RNAi. Results showed the ferritin mRNA was significantly down-regulated by 66.11% after RNAi. Western blot result showed that the expression level of ferritin protein was also depressed post RNAi. The method developed in this study proved to be reliable and effective for RNAi assay on marine bivalve cells. It would be an efficient tool for gene function analysis in marine bivalves and more studies based on primary cells of marine bivalves can be expected.
关键词	Marine Bivalve Primary Cell Culture Rnai Assay Gene Function Analysis
学科领域	Biochemistry & Molecular Biology
DOI	10.1007/s11033-012-1668-y
URL	查看原文
收录类别	SCI
语种	英语
WOS记录号	WOS:000305749200041
引用统计	被引频次：14[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/12375
专题	实验海洋生物学重点实验室
通讯作者	Liu, BZ (reprint author), Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China.
作者单位	1.Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
第一作者单位	中国科学院海洋研究所
推荐引用方式 GB/T 7714	You, Yanan,Huan, Pin,Liu, Baozhong,et al. RNAi assay in primary cells: a new method for gene function analysis in marine bivalve[J]. MOLECULAR BIOLOGY REPORTS,2012,39(8):8209-8216.
APA	You, Yanan,Huan, Pin,Liu, Baozhong,&Liu, BZ .(2012).RNAi assay in primary cells: a new method for gene function analysis in marine bivalve.MOLECULAR BIOLOGY REPORTS,39(8),8209-8216.
MLA	You, Yanan,et al."RNAi assay in primary cells: a new method for gene function analysis in marine bivalve".MOLECULAR BIOLOGY REPORTS 39.8(2012):8209-8216.

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