( Retroviral l...)Functional Cloning Using,,,ViraPort Retroviral cDNA Expression Libraries

Retroviral libraries with high-titer efficiency and larger insert potential

Functional Cloning Using
ViraPort Retroviral cDNA Expression Libraries

Katherine Felts Kim Zaharee Latha Sundar Jamie Limjoco
Anna Waesche Peter Vaillancourt
Stratagene

We introduce a set of human cDNA expression libraries inserted into the high-titer retroviral vector pFB. ViraPort retroviral libraries allow complex cDNA libraries to be efficiently introduced into virtually any mitotic cell type for screening based on gene function. The cDNA copy number per cell can be easily controlled by adjusting the multiplicity of infection (MOI). In this way, cell populations may be generated in which greater than 90% of infected cells contain one-to-five cDNAs. We describe the isolation of two known oncogenes and one cell-surface receptor from one of the libraries (Burkitts Lymphoma).

Table 1: Examples of Genes Cloned by Expression Screening

Gene Class

Selection/Screen

Cell-surface proteins

FACS sorting

Extracellular receptors

FACS sorting

Proliferation or survival

Growth factors

Factor-dependent growth

Oncogenes

Loss of contact-inhibition

Cell-cycle proteins

Loss of contact-inhibition

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