scholarly journals Creation and validation of a ligation-independent cloning (LIC) retroviral vector for stable gene transduction in mammalian cells

2012 ◽  
Vol 12 (1) ◽  
pp. 3 ◽  
Author(s):  
Asmita Patel ◽  
Anisleidys Muñoz ◽  
Katherine Halvorsen ◽  
Priyamvada Rai
Gene ◽  
1986 ◽  
Vol 45 (2) ◽  
pp. 121-129 ◽  
Author(s):  
Tony F. Weighous ◽  
James C. Cornette ◽  
Satish K. Sharma ◽  
W.Gary Tarpley

2001 ◽  
Vol 75 (11) ◽  
pp. 4973-4983 ◽  
Author(s):  
Eugene V. Barsov ◽  
William S. Payne ◽  
Stephen H. Hughes

ABSTRACT We have designed and characterized two new replication-competent avian sarcoma/leukosis virus-based retroviral vectors with amphotropic and ecotropic host ranges. The amphotropic vector RCASBP-M2C(797-8), was obtained by passaging the chimeric retroviral vector RCASBP-M2C(4070A) (6) in chicken embryos. The ecotropic vector, RCASBP(Eco), was created by replacing theenv-coding region in the retroviral vector RCASBP(A) with the env region from an ecotropic murine leukemia virus. It replicates efficiently in avian DFJ8 cells that express murine ecotropic receptor. For both vectors, permanent cell lines that produce viral stocks with titers of about 5 × 106 CFU/ml on mammalian cells can be easily established by passaging transfected avian cells. Some chimeric viruses, for example, RCASBP(Eco), replicate efficiently without modifications. For those chimeric viruses that do require modification, adaptation by passage in vitro or in vivo is a general strategy. This strategy has been used to prepare vectors with altered host range and could potentially be used to develop vectors that would be useful for targeted gene delivery.


2004 ◽  
Vol 324 (4) ◽  
pp. 1173-1178 ◽  
Author(s):  
Xiao-dong Liu ◽  
Shum-mei Ma ◽  
Yang Liu ◽  
Shu-zheng Liu ◽  
Alec Sehon

Biomaterials ◽  
2008 ◽  
Vol 29 (9) ◽  
pp. 1242-1249 ◽  
Author(s):  
Stephan Vogt ◽  
Peter Ueblacker ◽  
Christopher Geis ◽  
Bettina Wagner ◽  
Gabriele Wexel ◽  
...  

1996 ◽  
Vol 70 (1) ◽  
pp. 62-67 ◽  
Author(s):  
W Paulus ◽  
I Baur ◽  
F M Boyce ◽  
X O Breakefield ◽  
S A Reeves

2007 ◽  
Vol 35 (3) ◽  
pp. 275-282 ◽  
Author(s):  
Gro Leite Størvold ◽  
Elisabet Gjernes ◽  
Hanne Arenberg Askautrud ◽  
Anne-Lise Børresen-Dale ◽  
Charles M. Perou ◽  
...  

Blood ◽  
1998 ◽  
Vol 92 (9) ◽  
pp. 3163-3171 ◽  
Author(s):  
Andrea J. Schilz ◽  
Gaby Brouns ◽  
Heike Knöβ ◽  
Oliver G. Ottmann ◽  
Dieter Hoelzer ◽  
...  

Abstract Stable gene transfer to human pluripotent hematopoietic stem cells (PHSCs) is an attractive strategy for the curative treatment of many genetic hematologic disorders. In clinical trials, the levels of gene transfer to this cell population have generally been low, reflecting deficiencies in both the vector systems and transduction conditions. In this study, we have used a pseudotyped murine retroviral vector to transduce human CD34+ cells purified from bone marrow (BM) and umbilical cord blood (CB) under optimized conditions. After transduction, 71% to 97% of the hematopoietic cells were found to express a low-affinity nerve growth factor receptor (LNGFR) marker gene. Six weeks after transplantation into immunodeficient NOD/LtSz-scid/scid (NOD/SCID) mice, LNGFR expression was detected in 6% to 57% of CD45+ cells in eight of nine engrafted animals. Moreover, proviral DNA was detected in 8.3% to 45% of secondary colonies derived from BM cells of engrafted NOD/SCID mice. Our data show consistent transduction of SCID-repopulating cells (SRCs) and suggest that the efficiency of gene transfer to human hematopoietic repopulating cells can be improved using existing retroviral vector systems and carefully optimized transduction conditions. © 1998 by The American Society of Hematology.


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