RNA干扰abcg-5基因对犬弓首蛔虫感染性的影响

doi:10.11843/j.issn.0366-6964.2019.02.017

Abstract

Abstract:

This experiment was conducted to study the effect of ATP-binding cassette G5 gene of Toxocara canis. Specific siRNAs of the Tc-abcg-5 gene fragment were synthesized in vitro, and adult T. canis and eggs were interfered by using RNAi techniques. Results were as follows:Compared with siRNA-1020 group and siRNA-432 group, the transcriptional level of Tc-abcg-5 in siRNA-744 group was significantly reduced after 24 and 48 h's treatments, indicating that siRNA-744 had higher silencing efficiency. The delivery efficiency of siRNA-744 to eggs was compared by electroporation and immersion methods. The results showed that immersion method was more efficient. Interference of T. canis infective eggs were performed using siRNA-744 by soaking. Pathology study was used to analysis of mouse liver, brain and lung tissues on the 7th day after oral administration of interfered eggs, which showed that RNAi reduced the infectivity of T. canis infective eggs, thereby reducing the lung and liver lesions. It is suggested that the Tc-abcg-5 gene may participate in the growth and development of T. canis by affecting the development of eggs.

CLC Number:

S852.731

LUO Yongli, JIANG Aiyun, HU Zhigang, LI Xianglan, YANG Xiaodi, KUANG Ceyan, ZHOU Rongqiong. Effect of RNA Interference abcg-5 Gene on the Infectivity of Toxocara canis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(2): 390-397.

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References

[1] VENEGAS V, ZHOU Z. Two alternative mechanisms that regulate the presentation of apoptotic cell engulfment signal in Caenorhabditis elegans[J]. Mol Biol Cell, 2007, 18(8):3180-3192.
[2] ZHAO Z Y, SHEPS J A, LING V, et al. Expression analysis of ABC transporters reveals differential functions of tandemly duplicated genes in Caenorhabditis elegans[J]. J Mol Biol, 2004, 344(2):409-417.
[3] ARDELLI B F. Transport proteins of the ABC systems superfamily and their role in drug action and resistance in nematodes[J]. Parasitol Int, 2013, 62(6):639-646.
[4] ARDELLI B F, STITT L E, TOMPKINS J B. Inventory and analysis of ATP-binding cassette (ABC) systems in Brugia malayi[J]. Parasitology, 2010, 137(8):1195-1212.
[5] TRAN P N, BROWN S H, MITCHELL T W, et al. A female gametocyte-specific ABC transporter plays a role in lipid metabolism in the malaria parasite[J]. Nat Commun, 2014, 5:4773.
[6] ELBASHIR S M, HARBORTH J, LENDECKEL W, et al. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells[J]. Nature, 2001, 411(6836):494-498.
[7] MCCAFFREY A P, MEUSE L, PHAM T T, et al. RNA interference in adult mice[J]. Nature, 2002, 418(6893):38-39.
[8] KIM M H, YUAN X, OKUMURA S, et al. Successful inactivation of endogenous Oct-3/4 and c-mos genes in mouse preimplantation embryos and oocytes using short interfering RNAs[J]. Biochem Biophys Res Commun, 2002, 296(5):1372-1377.
[9] ALLEN R S, MILLGATE A G, CHITTY J A, et al. RNAi-mediated replacement of morphine with the nonnarcotic alkaloid reticuline in opium poppy[J]. Nat Biotechnol, 2004, 22(12):1559-1566.
[10] DUBOUZET J G, MORISHIGE T, FUJⅡ N, et al. Transient RNA silencing of scoulerine 9-O-methyltransferase expression by double stranded RNA in Coptis japonica protoplasts[J]. Biosci, Biotechnol, Biochem, 2005, 69(1):63-70.
[11] AL-ANOUTI F, ANANVORANICH S. Comparative analysis of antisense RNA, double-stranded RNA, and delta ribozyme-mediated gene regulation in Toxoplasma gondii[J]. Antisense Nucleic Acid Drug Dev, 2002, 12(4):275-281.
[12] OCÁDIZ-RUIZ R, FONSECA W, MARTÍNEZ M B, et al. Effect of the silencing of the Ehcp112 gene on the in vitro virulence of Entamoeba histolytica[J]. Parasit Vectors, 2013, 6(1):248-256.
[13] 王树奇,徐斌,刘建,等. 日本血吸虫醛糖还原酶基因RNA干扰效应的初步研究[J]. 中国寄生虫学与寄生虫病杂志, 2014, 32(1):17-21.
WANG S Q, XU B, LIU J, et al. Effect of RNA interference targeting Schistosoma japonicum aldose reductase gene[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2014, 32(1):17-21. (in Chinese)
[14] WANG X, CHEN W, TIAN Y, et al. RNAi-mediated silencing of enolase confirms its biological importance in Clonorchis sinensis[J]. Parasitol Res, 2014, 113(4):1451-1458.
[15] ZAWADZKI J L, PRESIDENTE P J, MEEUSEN E N, et al. RNAi in Haemonchus contortus:a potential method for target validation[J]. Trends Parasitol, 2006, 22(11):495-499.
[16] CHEN N, XU M J, NISBET A J, et al. Ascaris suum:RNAi mediated silencing of enolase gene expression in infective larvae[J]. Exp Parasitol, 2011, 127(1):142-146.
[17] DINH P T, BROWN C R, ELLING A A. RNA interference of effector gene Mc16D10L confers resistance against Meloidogyne chitwoodi in Arabidopsis and potato[J]. Phytopathology, 2014, 104(10):1098-1106.
[18] TIMMONS L, COURT D L, FIRE A. Ingestion of bacterially expressed dsRNAs can produce specific and potent genetic interference in Caenorhabditis elegans[J]. Gene, 2001, 263(1-2):103-112.
[19] WANG J, BARR M M. RNA interference in Caenorhabditis elegans[J]. Methods Enzymol, 2005, 392:36-55.
[20] SELKIRK M E, HUANG S C, KNOX D P, et al. The development of RNA interference (RNAi) in gastrointestinal nematodes[J]. Parasitology, 2012, 139(5):605-612.
[21] 张媛媛,许蓉,何川川,等. 日本血吸虫ELAV-like 2基因的克隆、表达及功能分析[J]. 畜牧兽医学报, 2018, 49(9):1987-1997.
ZHANG Y Y, XU R, HE C C, et al. Cloning, expression and functional analysis of ELAV-like 2 gene of Schistosoma japonicum[J]. Acta Veterinaria et Zootechnica Sinica, 2018, 49(9):1987-1997. (in Chinese)
[22] LUO Y L, MA G X, LUO Y F, et al. Tissue expression pattern of ABCG transporter indicates functional roles in reproduction of Toxocara canis[J]. Parasitol Res, 2018, 117(3):775-782.
[23] GREENBERG R M. Schistosome ABC multidrug transporters:from pharmacology to physiology[J]. Int J Parasitol Drugs Drug Resist, 2014, 4(3):301-309.
[24] KASINATHAN R S, MORGAN W M, GREENBERG R M. Genetic knockdown and pharmacological inhibition of parasite multidrug resistance transporters disrupts egg production in Schistosoma mansoni[J]. PLoS Negl Trop Dis, 2011, 5(12):e1425-e1433.

Effect of RNA Interference abcg-5 Gene on the Infectivity of Toxocara canis

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