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Abstract  Lentiviral technology is a powerful tool for the creation of stable transgenic animals. However, uncertainties have remained whether constitutive promoters resist long-term silencing. We used concentrated HIV-1 based lentiviral vectors to create stable transgenic BALB/c mice by perivitelline injection. In our vectors eGFP expression was driven by the human EF1α promoter. The established transgenic animals were analyzed for eGFP expression by in vivo fluorescence imaging, PCR, histology and flow-cytometry. eGFP expression showed even distribution without mosaicism; however, tissue-dependent differences of eGFP expression were observed. Up to the sixth generation only one newborn showed eGFP inactivation. eGFP  transgenic bone marrow cells efficiently provided long-term haemopoietic repopulation in radiation chimeras, regenerating all bone marrow-derived lineages with eGFP  cells with distinct eGFP expression profiles. The established eGFP  BALB/c mouse strain is expected to be extremely useful in various immunological experiments. [ABSTRACT FROM AUTHOR]

Abstract  Lentiviral vectors are now recognised as an efficient transgene delivery system which can result in greater than 90% of founder animals carrying the transgene. Vector injection into the perivitelline space has emerged as the standard delivery method but is limited by the need for high-titre lentivirus vector preparations. Based on a modified perivitelline injection method we demonstrate that transgenic animals can be generated from low-titre virus vector preparations further simplifying lentiviral transgenesis. Repeat injection of 107 TU/ml vector preparation resulted in 23% of embryos carrying the transgene compare to 1% from a single injection. Embryos exposed to repeat injection of vector developed to blastocyst with the same efficiency as non-injected embryos and produced transgenic mice capable of transmitting the transgene through the germline [ABSTRACT FROM AUTHOR]

Summary: We have developed an online tool called mirEval which can search sequences of up to 10 000 nt for novel microRNAs in multiple organisms. It is a comprehensive tool, easy to use and very informative. It will allow users with no prior knowledge of in-silico detection of microRNAs to take advantage of the most successful approaches to investigate sequences of interest. Availability: The mirEval web server is available at http://tagc.univ-mrs.fr/mireval Contact: W.Ritchie@centenary.org.au Supplementary information: Supplementary data are available at Bioinformatics online. [ABSTRACT FROM AUTHOR]