A retroviral vector suitable for ultrasound image-guided gene delivery to mouse brain

Abstract

Gene transfer to the early-stage embryonic brain using the ultrasound image-guided gene delivery (UIGD) technique has proven to be valuable for investigating brain development. Thus far, this technology has been restricted to the study of embryonic neurogenesis. When this technique is designed to be employed for the study in adult animals, a long-term stable gene expression will be required. We attempted to develop a retroviral vector suitable for expressing exogenous genes in the brains of postnatal and adult mice in the context of the UIGD technique. Retroviral vectors containing four different long terminal repeats (LTRs) (each from Moloney murine leukemia virus (MoMLV), murine stem cell virus (MSCV), myeloproliferative sarcoma virus (MPSV) and spleen focus-forming virus (SFFV)) were compared using the well-known CE vector having the EF1a internal promoter as a control. The MS vector containing MSCV LTR produced a higher viral titer and a higher level of gene expression than other vectors including CE. The MS vector drove the gene expression in cultured neural stem cells for 3 weeks. Furthermore, the MS vector could efficiently deliver the gene to the mouse central nervous system, as transgene expression was found in various regions of the brains and spinal cords as well as in all major neural cell types. The data from an in vivo luciferase imaging analysis showed that the gene expression from the MS vector was sustainable for almost 3 months. Our data suggested that the MS vector would be suitable to construct mice containing the transgene expressed in the brain or spinal cord in a quick and cost-effective manner.

Quaking-5 suppresses aggressiveness of lung cancer cells through inhibiting β-catenin signaling pathway

“…Then, QKI-5 was subcloned into pMSCV-puro plasmid. The control (empty vector) or QKI-5-expressing retrovirus was constructed as previously described [ 25 ]. The infection of retroviral particles was performed as previously described [ 18 ].…”

Upregulation of Nrf2 expression by human cytomegalovirus infection protects host cells from oxidative stress

“…The PCR product was cloned into the BamHI site of the retroviral vector MSIG (Jang et al, 2012). This vector contains the LTR originally from a murine stem cell virus, and eGFP is expressed as a bicistronic message using the internal ribosome entry site (IRES) from encephalomyocarditis virus.…”

GSK3β, But Not GSK3α, Inhibits the Neuronal Differentiation of Neural Progenitor Cells As a Downstream Target of Mammalian Target of Rapamycin Complex1

“…The MS vector containing the long terminal repeat from a murine stem cell virus was described previously [18]. The mouse GSK3a sequence was synthesized (Bioneer), and the mouse GSK3b sequence was amplified from Raw 264.7 cDNA using primers listed in Table 1.…”