Reprogramming of adult cells to generate induced pluripotent stem cells (iPS cells) has opened new therapeutic opportunities; however, little is known about the possibility of in vivo reprogramming within tissues. Here we show that transitory induction of the four factors Oct4, Sox2, Klf4 and c-Myc in mice results in teratomas emerging from multiple organs, implying that full reprogramming can occur in vivo. Analyses of the stomach, intestine, pancreas and kidney reveal groups of dedifferentiated cells that express the pluripotency marker NANOG, indicative of in situ reprogramming. By bone marrow transplantation, we demonstrate that haematopoietic cells can also be reprogrammed in vivo. Notably, reprogrammable mice present circulating iPS cells in the blood and, at the transcriptome level, these in vivo generated iPS cells are closer to embryonic stem cells (ES cells) than standard in vitro generated iPS cells. Moreover, in vivo iPS cells efficiently contribute to the trophectoderm lineage, suggesting that they achieve a more plastic or primitive state than ES cells. Finally, intraperitoneal injection of in vivo iPS cells generates embryo-like structures that express embryonic and extraembryonic markers. We conclude that reprogramming in vivo is feasible and confers totipotency features absent in standard iPS or ES cells. These discoveries could be relevant for future applications of reprogramming in regenerative medicine.

A novel DNA polymerase has been identified in human cells. Human DNA polymerase mu (Pol mu), consisting of 494 amino acids, has 41% identity to terminal deoxynucleotidyltransferase (TdT). Human Pol mu, overproduced in Escherichia coli in a soluble form and purified to homogeneity, displays intrinsic terminal deoxynucleotidyltransferase activity and a strong preference for activating Mn(2+) ions. Interestingly, unlike TdT, the catalytic efficiency of polymerization carried out by Pol mu was enhanced by the presence of a template strand. Using activating Mg(2+) ions, template-enhanced polymerization was also template-directed, leading to the preferred insertion of complementary nucleotides, although with low discrimination values. In the presence of Mn(2+) ions, template-enhanced polymerization produced a random insertion of nucleotides. Northern-blotting and in situ analysis showed a preferential expression of Pol mu mRNA in peripheral lymphoid tissues. Moreover, a large proportion of the human expressed sequence tags corresponding to Pol mu, present in the databases, derived from germinal center B cells. Therefore, Pol mu is a good candidate to be the mutator polymerase responsible for somatic hyper- mutation of immunoglobulin genes.

Rap1 is a component of the shelterin complex at mammalian telomeres, but its in vivo role in telomere biology has remained largely unknown to date. Here we show that Rap1 deficiency is dispensable for telomere capping but leads to increased telomere recombination and fragility. We generated cells and mice deleted for Rap1; mice with Rap1 deletion in stratified epithelia were viable but had shorter telomeres and developed skin hyperpigmentation in adulthood. By performing chromatin immunoprecipitation coupled with ultrahigh-throughput sequencing, we found that Rap1 binds to both telomeres and to extratelomeric sites through the (TTAGGG)(2) consensus motif. Extratelomeric Rap1-binding sites were enriched at subtelomeric regions, in agreement with preferential deregulation of subtelomeric genes in Rap1-deficient cells. More than 70% of extratelomeric Rap1-binding sites were in the vicinity of genes, and 31% of the genes deregulated in Rap1-null cells contained Rap1-binding sites, suggesting a role for Rap1 in transcriptional control. These findings place a telomere protein at the interface between telomere function and transcriptional regulation.

Seasonally dry tropical forests are distributed across Latin America and the Caribbean and are highly threatened, with less than 10% of their original extent remaining in many countries. Using 835 inventories covering 4660 species of woody plants, we show marked floristic turnover among inventories and regions, which may be higher than in other neotropical biomes, such as savanna. Such high floristic turnover indicates that numerous conservation areas across many countries will be needed to protect the full diversity of tropical dry forests. Our results provide a scientific framework within which national decision-makers can contextualize the floristic significance of their dry forest at a regional and continental scale. N eotropical seasonally dry forest (dry forest) is a biome with a wide and fragmented distribution, found from Mexico to Argentina and throughout the Caribbean (1, 2) ( Fig. 1). It is one of the most threatened tropical forests in the world (3), with less than 10% of its original extent remaining in many countries (4).Following other authors (5, 6), we define dry forest as having a closed canopy, distinguishing it from more open, grass-rich savanna. It occurs on fertile soils where the rainfall is less thañ 1800 mm per year, with a period of 3 to 6 months receiving less than 100 mm per month (5-7), during which the vegetation is mostly deciduous. Seasonally dry areas, especially in Peru and Mexico, were home to pre-Columbian civilizations, so human interaction with dry forest has a long history (8). The climates and fertile soils of dry forest regions have led to higher human population densities and an increasing demand for energy and land, enhancing degradation (9). More recently, destruction of dry forest has been accelerated by intensive cultivation of crops, such as sugar cane, rice and soy, or by conversion to pasture for cattle.Dry forest is in a critical state because so little of it is intact, and of the remnant areas, little is protected (3). For example, only 1.2% of the total Caatinga region of dry forest in Brazil is fully protected compared with 9.9% of the Brazilian Amazon (10). Conservation actions are urgently needed to protect dry forest's unique biodiversity-many plant species and even genera are restricted to it and reflect an evolutionary history confined to this biome (1).We evaluate the floristic relationships of the disjunct areas of neotropical dry forest and highlight those that contain the highest diversity and endemism of woody plant species. We also explore woody plant species turnover across geographic space among dry forests. Our results provide a framework to allow the conservation significance of each separate major region of dry forest to be assessed at a continental scale. Our analyses are based on a subset of a data set of 1602 inventories made in dry forest and related semi-deciduous forests from Mexico and the Caribbean to Argentina and Paraguay that covers 6958 woody species, which has been compiled by the Latin American and Caribbean Seasonally Dry Tropica...