shRNA-Induced Gene Knockdown In Vivo to Investigate Neutrophil Function.
To silence genes in neutrophils efficiently, we exploited the RNA interference and developed an shRNA-based gene knockdown technique. This method involves transfection of mouse bone marrow-derived hematopoietic stem cells with retroviral vector carrying shRNA directed at a specific gene. Transfected stem cells are then transplanted into irradiated wild-type mice. After engraftment of stem cells, the transplanted mice have two sets of circulating neutrophils. One set has a gene of interest knocked down while the other set has full complement of expressed genes. This efficient technique provides a unique way to directly compare the response of neutrophils with a knocked-down gene to that of neutrophils with the full complement of expressed genes in the same environment.
Department of Pharmacology, Vascular Biology and Therapeutic Program, Yale University School of Medicine, 10 Amistad, P.O. Box 208089, New Haven, CT, 06520, USA.
Abdul
A
Basit
Department of Pharmacology, Vascular Biology and Therapeutic Program, Yale University School of Medicine, 10 Amistad, P.O. Box 208089, New Haven, CT, 06520, USA.
Wenwen
W
Tang
Department of Pharmacology, Vascular Biology and Therapeutic Program, Yale University School of Medicine, 10 Amistad, P.O. Box 208089, New Haven, CT, 06520, USA. dianqing.wu@yale.edu.
Dianqing
D
Wu
D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism; D016026:Bone Marrow Transplantation; D002460:Cell Line; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D005822:Genetic Vectors / Q000235:genetics; D051379:Mice; D009504:Neutrophils / Q000502:physiology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic
D034741:RNA, Small Interfering
Gene knockdown; Neutrophils; RNA interference; Retroviral vectors; Short hairpin RNA; Transplantation