Construction of lncRNA4.9 interfering lentivirus vector
10.3760/cma.j.issn.1003-9279.2020.01.017
- VernacularTitle:lncRNA4.9干扰慢病毒载体的构建
- Author:
Lifang LIU
1
;
Wei LI
;
Ran TAO
;
Huamei LI
;
Luyan CHEN
;
Shiqiang SHANG
Author Information
1. 浙江大学医学院附属儿童医院
- Keywords:
Human cytomegalovirus;
Long non-coding RNA;
Interfering lentivirus vector
- From:
Chinese Journal of Experimental and Clinical Virology
2020;34(1):83-86
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct an interfering lentivirus vector of long non-coding RNA 4.9 (lncRNA4.9) transcribed by human cytomegalovirus (HCMV).Methods Three interfering sequences targeting lncRNA 4.9 were designed and synthesized.The shuttle plasmid GV248 and the target interfering sequence were combined and constructed.The recombinant plasmid was co-transfected with the skeleton plasmids pHelper1.0 and pHelper2.0 to 293T cells.Viral particles were collected and copies were determined.The interfering lentivirus vector was transfected into THP-1 cells to observe the fluorescence expression,and the interfering efficiency was detected by real-time RT-PCR.Results Three groups of lentivirus interference vectors (LV1,LV2,LV3) were constructed,LV2 and LV3 can interfere with the expression of lncRNA4.9,and the interference efficiency of LV2 group was the highest.Conclusions The interfering lentivirus vector of lncRNA 4.9 was successfully constructed,which laid a foundation for further study on the function of lncRNA 4.9.
