Rapid assembly of customized TALENs into multiple

Transcriptional activator-like effector nucleases (TALENs) have become a powerful tool for genome editing. Here we present an efficient TALEN assembly approach in which TALENs are assembled by direct Golden Gate ligation into Gateway® Entry vectors from a repeat variable di-residue (RVD) plasmid array. We constructed TALEN pairs targeted to mouse Ddx3 subfamily genes, and demonstrated that our modified TALEN assembly approach efficiently generates accurate TALEN moieties that effectively introduce mutations into target genes. We generated "user friendly" TALEN Entry vectors containing TALEN expression cassettes with fluorescent reporter genes that can be efficiently transferred via Gateway (LR) recombination into different delivery systems. We demonstrated that the TALEN Entry vectors can be easily transferred to an adenoviral delivery system to expand application to cells that are difficult to transfect. Since TALENs work in pairs, we also generated a TALEN Entry vector set that combines a TALEN pair into one PiggyBac transposon-based destination vector. The approach described here can also be modified for construction of TALE transcriptional activators, repressors or other functional domains. © 2013 Zhang et al

The Iranian education and academic mobility model

The paper discusses the evolution of the educational model and trends of academic mobility in the Islamic Republic of Iran (IRI). It analyzes the socio-demographic structure of the population, the dynamics of the number of young people, as well as the dynamics and composition of the country’s school in the country. There is also an examination of the ways in which the process of emigration from Iran had evolved in the context of the country’s socio-political history of the state in the 20th-21st centuries. Various levels of the Iranian education system are considered. The author examines the internationalization trends in higher education. The forms and trends of academic mobility, including exit and entry mobility, are delineated. In recent years, the visiting academic mobility of Iranian students has been targeted at the U.S., Turkey, Italy, Canada, and the UAE. There is a contemplation of the methods used for selecting Iranian students for studying abroad at the state budget’s expense, which is based on a system of state quotas and fairly high requirements for applicants. Iranian students studying abroad receive an academic scholarship. Those who receive such a scholarship must pay a deposit before being issued a final permit, and commit to returning to Iran after graduation and working for up to six years. Recently, increasing the number of foreign students in Iran has become one of the priorities of the Ministry of Science, Research and Technology. The admission of foreign students has presently become one of the most important tasks undertaken by universities. One way to attract more foreign students and compete for them is to raise the status of universities and the country. Iran is growing more attractive as a recipient of international students from certain neighboring countries (Afghanistan, Iraq, Syria, Lebanon, Pakistan, and China). The role of academic mobility in the integration of Iran into the international educational and scientific space and the contribution of academic exchanges to the development of the national culture of Iran are emphasized. © 2019, CA and C Press AB. All rights reserved

Epigenetic resetting of human pluripotency

Much attention has focussed on the conversion of human pluripotent stem cells (PSCs) to a more naïve developmental status. Here we provide a method for resetting via transient histone deacetylase inhibition. The protocol is effective across multiple PSC lines and can proceed without karyotype change. Reset cells can be expanded without feeders with a doubling time of around 24 h. WNT inhibition stabilises the resetting process. The transcriptome of reset cells diverges markedly from that of primed PSCs and shares features with human inner cell mass (ICM). Reset cells activate expression of primate-specific transposable elements. DNA methylation is globally reduced to a level equivalent to that in the ICM and is non-random, with gain of methylation at specific loci. Methylation imprints are mostly lost, however. Reset cells can be re-primed to undergo tri-lineage differentiation and germline specification. In female reset cells, appearance of biallelic X-linked gene transcription indicates reactivation of the silenced X chromosome. On reconversion to primed status, XIST-induced silencing restores monoallelic gene expression. The facile and robust conversion routine with accompanying data resources will enable widespread utilisation, interrogation, and refinement of candidate naïve cells