title: [Screening lentiviral vectors carrying effective siRNA of the ROCK2 gene].
abstract: OBJECTIVE
  To screen the lentiviral vector carrying siRNA and capable of significantly suppressing the ROCK2 gene expression in the corpus cavernosum smooth muscle cells of spontaneously hypertensive rats (SHR).
  
  
  METHODS
  We designed and synthesized 4 siRNA fragments targeting the ROCK2 gene and packaged them into lentiviral vectors. We collected corpus cavernosum smooth muscle cell samples from 5 male SHRs and randomly divided them into groups A (non-transfection control), B (GFP lentiviral transfection), C, D, E, and F (lentiviral transfection with siRNA fragments 1 -4 targeting the ROCK2 gene). Each group consisted of 5 samples and each sample 3 x 10(4) cells. At 48 hours after transfecting MOI = 80 into the SHR corpus cavernosum smooth muscle cells, we detected the expression of GFP under the fluorescent microscope and the mRNA expression of the ROCK2 gene by RT-PCR.
  
  
  RESULTS
  The transfection efficiency of the SHR corpus cavernosum smooth muscle cells was > 50%. Compared with group A, the expression of ROCK2 mRNA in the corpus cavernosum smooth muscle cells showed no remarkable change in group B (P > 0.05) but was inhibited very significantly in C ([43.91 +/- 8.19]%), D ([47.15 +/- 6.64]%), and F ([25.7 +/- 6.03]%) (P < 0.01), and significantly in E ([16.81 +/- 5.94]%, P < 0.05).
  
  
  CONCLUSIONS
  We successfully constructed 4 lentiviral vectors carrying siRNA targeting the ROCK2 gene, all of which can significantly suppress the ROCK2 expression in the SHR corpus cavernosum smooth muscle cells, and one has a highly strong inhibitory effect.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Peng
authors.forename: Fu-Nian
authors.initials: FN
authors.lastname: Ma
authors.forename: Rui
authors.initials: R
authors.lastname: Jiang
authors.forename: Feng
authors.initials: F
authors.lastname: Chen
mesh_terms: D000818:Animals; D020868:Gene Silencing; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics*; D008297:Male; D032389:Myocytes, Smooth Muscle; D010413:Penis / Q000166:cytology; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering; D011897:Random Allocation; D051381:Rats; D011918:Rats, Inbred SHR; D014162:Transfection / Q000379:methods*; D054460:rho-Associated Kinases / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; C580818:ROCK2 protein, rat; D054460:rho-Associated Kinases