Transgenic mouse models of the truncated platelet integrin β3 cytoplasmic tail established by stem cell transplantation.
10.7534/j.issn.1009-2137.2013.03.026
- Author:
Xiong-Ying CUI
1
;
Xiao-Feng SHI
;
Jian-Song HUANG
;
Ping LIU
;
Lan-Lan TAO
;
Yu-Lan ZHOU
;
Zheng RUAN
;
Xiao-Dong XI
Author Information
1. Shanghai Jiaotong University School of Medicine, Shanghai, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Blood Platelets;
metabolism;
Genetic Vectors;
Hematopoietic Stem Cell Transplantation;
Integrin beta3;
metabolism;
Mice;
Mice, Inbred C57BL;
Mice, Transgenic;
Retroviridae;
genetics
- From:
Journal of Experimental Hematology
2013;21(3):667-673
- CountryChina
- Language:Chinese
-
Abstract:
This study was purpose to establish the transgenic mouse models of the truncated platelet integrin β3 by retrovirus-infected hematopoietic stem cells (HSCs) transplantation and to provide the basis for further study of the role of integrin β3 cytoplasmic domain in platelet bi-directional signaling pathways. Wild-type β3, β3-Δ759 (R(760) GT(762) truncated β3) and β3-Δ754 (T(755) NITYRGT(762) truncated β3) cDNAs were subcloned into MSCV MigR1 retroviral vector bearing a GFP gene and packaged into infective retrovirus with BOSC23 cell strain. The bone marrow HSCs of the β3 deficient mice were infected by the retroviruses, and transplanted into lethally-irradiated wild type C57BL/6 mice. GFP positive rate and surface β3 expression of the recipients' platelets at 6 to 8 weeks after transplantation were detected by flow cytometry to evaluate the transgenic efficiency. The results showed that four kinds of transgenic mouse models including vector, wild-type β3, β3-Δ759 and β3-Δ754 were established successfully. GFP positive rates of transgenic mouse platelets ranged from 18% to 66% and the β3 expression of transgenic mouse reached heterozygote (β3(+/-) level of mouse). It is concluded that establishment of transgenic mouse models mediated by retrovirus-infected HSCs transplantation is a feasible, fast, and high throughput transgenic approach and laid a solid foundation for further research on the role of integrin β3 cytoplasmic domain for bi-directional signaling of platelets in vivo, and for the gene therapy of platelet disorders.
