title: [Preparation and knockdown efficiency evaluation of shRNA expressing lentiviral vector targeting B7-H1].
abstract: OBJECTIVE
  To design and package shRNA expressing lentiviral particles targeting B7-H1, and evaluate their inhibitory effect on B7-H1 expression in U251 cells.
  
  
  METHODS
  Three shRNAs targeting B7-H1 was designed and the sense and antisense primers were produced by chemical synthesis. After annealing, they were linked into restriction enzyme digested pLKO.1 vector. Confirmed by DNA sequencing, the lentiviral particles were packaged and applied to infect U251 cells. qRT-PCR and Western blotting were used to detect the B7-H1 mRNA and protein levels respectively.
  
  
  RESULTS
  qRT-PCR and Western blotting showed that two of the three shRNAs effectively knocked-down B7-H1 expression in U251 cells.
  
  
  CONCLUSIONS
  The packaged lentiviral particles can specifically inhibit B7-H1 expression, which will be helpful for further functional study on B7-H1.
authors: Department of Neurosurgery, Fourth Military Medical University, Xi'an, China.
authors: Luokai
authors: L
authors: Huangfu
authors: Xi
authors: X
authors: Wang
authors: Zhangyan
authors: Z
authors: Guo
authors: Wenjin
authors: W
authors: Xi
authors: Shengjia
authors: S
authors: Shi
authors: Fan
authors: F
authors: Yang
authors: Xiaoyan
authors: X
authors: Chen
authors: Angang
authors: A
authors: Yang
authors: Jianning
authors: J
authors: Zhang
authors: Weihong
authors: W
authors: Wen
mesh_terms: D000818:Animals; D060890:B7-H1 Antigen / Q000172:deficiency* / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D045744:Cell Line, Tumor; D004262:DNA Restriction Enzymes / Q000378:metabolism; D015870:Gene Expression; D055785:Gene Knockdown Techniques / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*
chemical_list: D060890:B7-H1 Antigen; D034741:RNA, Small Interfering; D004262:DNA Restriction Enzymes