Asynchronous combinatorial action of four regulatory factors activates Bcl11b for T cell commitment

During T cell development, multipotent progenitors relinquish competence for other fates and commit to the T cell lineage by turning on Bcl11b, which encodes a transcription factor. To clarify lineage commitment mechanisms, we followed developing T cells at the single-cell level using Bcl11b knock-in fluorescent reporter mice. Notch signaling and Notch-activated transcription factors collaborate to activate Bcl11b expression irrespectively of Notch-dependent proliferation. These inputs work via three distinct, asynchronous mechanisms: an early locus 'poising' function dependent on TCF-1 and GATA-3, a stochastic-permissivity function dependent on Notch signaling, and a separate amplitude-control function dependent on Runx1, a factor already present in multipotent progenitors. Despite their necessity for Bcl11b expression, these inputs act in a stage-specific manner, providing a multitiered mechanism for developmental gene regulation

Asynchronous combinatorial action of four regulatory factors activates Bcl11b for T cell commitment

During T cell development, multipotent progenitors relinquish competence for other fates and commit to the T cell lineage by turning on Bcl11b, which encodes a transcription factor. To clarify lineage commitment mechanisms, we followed developing T cells at the single-cell level using Bcl11b knock-in fluorescent reporter mice. Notch signaling and Notch-activated transcription factors collaborate to activate Bcl11b expression irrespectively of Notch-dependent proliferation. These inputs work via three distinct, asynchronous mechanisms: an early locus 'poising' function dependent on TCF-1 and GATA-3, a stochastic-permissivity function dependent on Notch signaling, and a separate amplitude-control function dependent on Runx1, a factor already present in multipotent progenitors. Despite their necessity for Bcl11b expression, these inputs act in a stage-specific manner, providing a multitiered mechanism for developmental gene regulation

Shipment of Mice and Embryos

Following the advent of transgenic technology and the subsequent establishment of international consortia for broad-based genomic analysis of the mouse, the need for sharing of resources in the exchange of mouse models and material has reached an all time high. Live animals, embryos, and gametes are ever more increasingly shipped around the world in an exchange or purchase of research material. In this chapter, we shall look at requirements for shipping of live animals locally, as well as review the current requirements for receipt of live animals by governing authorities in Europe, the UK, the USA, Australia, and China. We shall also review procedures for shipping of cryopreserved embryos and gametes, also embryos at ambient temperatures

Mammalian and Avian Transgenesis — New Approaches

Mammalian and Avian Transgenesis presents a collection of novel methods for the production of a wide range of transgenic animals. The manual focuses largely on mice, but also contains protocols for successful transgenesis in rats, cows, pigs and birds. The manual provides detailed, step-by-step protocols covering all aspects of the production of transgenic animals, including the use of lentiviral vectors in gene transfer, intracytoplasmic sperm injection, nuclear transfer, large insert transgenesis, conditional gene expression systems, the use of reporter genes in transgenesis and transgenesis in large animals and birds. The text is supplemented by superb color photos. While the focus is on newly established techniques, the fundamental methods of transgenesis are also covered for those new to the field. Thus this manual is perfectly suited for those wishing to adopt new technologies in transgenesis

Mammalian and Avian Transgenesis — New Approaches

Mammalian and Avian Transgenesis presents a collection of novel methods for the production of a wide range of transgenic animals. The manual focuses largely on mice, but also contains protocols for successful transgenesis in rats, cows, pigs and birds. The manual provides detailed, step-by-step protocols covering all aspects of the production of transgenic animals, including the use of lentiviral vectors in gene transfer, intracytoplasmic sperm injection, nuclear transfer, large insert transgenesis, conditional gene expression systems, the use of reporter genes in transgenesis and transgenesis in large animals and birds. The text is supplemented by superb color photos. While the focus is on newly established techniques, the fundamental methods of transgenesis are also covered for those new to the field. Thus this manual is perfectly suited for those wishing to adopt new technologies in transgenesis

Introducing Filipino wordnet

In this paper, we introduce the Filipino wordnet project (FilWordNet). Filipino is the national language of the Philippines spoken by some 90 million people as their first or second language. However, it has historically had a limited number of computational linguistics resources. Creating the Filipino wordnet can be seen as the first step to enable a wide range of research projects. We describe our process of building a wordnet, including issues with the Filipino language itself, its morphology and structure. © Global Wordnet Conference, GWC 2010. All rights reserved

Oct4 kinetics predict cell lineage patterning in the early mammalian embryo

Transcription factors are central to sustaining pluripotency, yet little is known about transcription factor dynamics in defining pluripotency in the early mammalian embryo. Here, we establish a fluorescence decay after photoactivation (FDAP) assay to quantitatively study the kinetic behaviour of Oct4, a key transcription factor controlling pre-implantation development in the mouse embryo. FDAP measurements reveal that each cell in a developing embryo shows one of two distinct Oct4 kinetics, before there are any morphologically distinguishable differences or outward signs of lineage patterning. The differences revealed by FDAP are due to differences in the accessibility of Oct4 to its DNA binding sites in the nucleus. Lineage tracing of the cells in the two distinct sub-populations demonstrates that the Oct4 kinetics predict lineages of the early embryo. Cells with slower Oct4 kinetics are more likely to give rise to the pluripotent cell lineage that contributes to the inner cell mass. Those with faster Oct4 kinetics contribute mostly to the extra-embryonic lineage. Our findings identify Oct4 kinetics, rather than differences in total transcription factor expression levels, as a predictive measure of developmental cell lineage patterning in the early mouse embryo