Role of a heterologous retroviral transport element in the development of genetic complementation assay for mouse mammary tumor virus (MMTV) replication.
The mouse mammary tumor virus (MMTV) is a type B retrovirus that is unique from other retroviruses in having multiple "tissue specific" and "hormone inducible" promoters. This unique feature has lead to the increasing interest in studying the biology of MMTV replication with the ultimate goal of developing MMTV based vectors for potentially targeted human gene therapy. In this report, we describe, for the first time, the establishment of an in vivo genetic complementation assay to study various aspects of MMTV replication. In the assay described here, the function of MMTV Rem/RmRE regulatory pathway has been successfully substituted by a heterologous retroviral constitutive transport element (CTE) from Mason Pfizer Monkey Virus (MPMV) for mature MMTV particle production. Our results revealed that in the absence of MPMV CTE or Rem/RmRE, RNA transcribed from MMTV Gag-Pol expression plasmids were efficiently transported to the cytoplasm. However, the presence of CTE was indispensable for Gag-Pol protein expression. In addition, we report the development of MMTV based vectors in which the packageable RNA was transcribed either from MMTV LTR or from a chimeric LTR, which could successfully be packaged and propagated by particles produced from MMTV Gag-Pol expression plasmids containing a heterologous transport element. The role of MPMV CTE in the transport of MMTV transfer vector RNA was not found to be significant. Development of such an assay should not only shed light on how MMTV regulates its gene expression, but also should provide additional molecular tools for delineating the packaging determinants for MMTV, which is imperative for the development of novel vectors for targeted and inducible gene therapy.
Rizvi
Tahir A
TA
Department of Microbiology & Immunology, Faculty of Medicine and Health Sciences (FMHS), United Arab Emirates University (UAEU), P.O. Box 17666, Al Ain, UAE. tarizvi@uaeu.ac.ae
Ali
Jahabar
J
Phillip
Pretty Susan
PS
Ghazawi
Akela
A
Jayanth
Preethi
P
Mustafa
Farah
F
D000818:Animals; D002460:Cell Line; D015685:Fusion Proteins, gag-pol / Q000378:metabolism; D015967:Gene Expression Regulation, Viral; D018014:Gene Transfer Techniques; D005814:Genes, Viral; D005816:Genetic Complementation Test / Q000379:methods*; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics / Q000502:physiology*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics*; D019065:Virus Assembly / Q000235:genetics; D014779:Virus Replication / Q000235:genetics
D015685:Fusion Proteins, gag-pol; D012367:RNA, Viral