Journal Article

Generation of RCAS Vectors Useful for Functional Genomic Analyses

Stacie K. Loftus, Denise M. Larson, Dawn Watkins-Chow, Deanna M. Church and William J. Pavan

in DNA Research

Published on behalf of Kazusa DNA Research Institute

Volume 8, issue 5, pages 221-226
Published in print January 2001 | ISSN: 1340-2838
Published online January 2001 | e-ISSN: 1756-1663 | DOI: http://dx.doi.org/10.1093/dnares/8.5.221

Show Summary Details

Preview

Avian leukosis type A virus-derived retroviral vectors have been used to introduce genes into cells expressing the corresponding avian receptor tv-a. This includes the use of Replication-Competent Avian sarcoma-leukosis virus (ASLV) long terminal repeat (LTR) with Splice acceptor (RCAS) vectors in the analysis of avian development, human and murine cell cultures, murine cell lineage studies and cancer biology. Previously, cloning of genes into this virus was difficult due to the large size of the vector and sparse cloning sites. To overcome some of the disadvantages of traditional cloning using the RCASBP-Y vector, we have modified the RCASBP-Y to incorporate “Gateway” site-specific recombination cloning of genes into the construct, either with or without HA epitope tags. We have found the repetitive “att” sequences, which are the targets for site-specific recombination, do not impair the production of infectious viral particles or the expression of the gene of interest. This is the first instance of site-specific recombination being used to generate retroviral gene constructs. These viral constructs will allow for the efficient transfer and expression of cDNAs needed for functional genomic analyses.

Keywords: RCAS; tv-a; retrovirus; recombination; Gateway; gene expression

Journal Article.  0 words. 

Subjects: Genetics and Genomics

Users without a subscription are not able to see the full content. Please, subscribe or login to access all content.