Gene Transfer in Avian Embryos Using Replication-Competent Retroviruses
A series of replication-competent, avian-specific retroviral vectors, known as RCAS or RCAN, were developed by Hughes et al. (1) and used successfully by a rapidly expanding number of groups to assess gene function directly in ovo (e.g.,(2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)). These proviral vectors are derived from the Rous sarcoma virus (RSV) and contain a unique ClaI restriction site in place of the region normally encoding the src oncogene, into which foreign DNA fragments of up to approximately 2.4 kb can be inserted. An E. coli plasmid backbone allows the gene of choice to be introduced by standard subcloning techniques, while retention of the viral long terminal repeat (LTR) sequences together with sequences encoding the viral gag, pol, and env genes facilitates viral replication and transmission. RCAN is a variant of RCAS, from which the splice acceptor immediately upstream of the ClaI site has been removed, preventing translation of the inserted gene,...
KeywordsLong Terminal Repeat Rous Sarcoma Virus Chicken Serum HEPES Buffer Saline Plastic Centrifuge Tube
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