Emerging technology to measure habitat quality and behavior of grouse: Examples from studies of greater sage-grouse

An increasing number of threats, both natural (e.g. fires, drought) and anthropogenic (e.g. agriculture, infrastructure development), are likely to affect both availability and quality of plants that grouse rely on for cover and food. As such, there is an increasing need to monitor plants and their use by grouse over space and time to better predict how changes in habitat quality influence the behavior of grouse. We use the greater sage-grouse Centrocercus urophasianus to showcase how new technology can be used to advance our understanding of the ecology, behavior and conservation of grouse. We demonstrate how laser, spectral and chemical detectors and unmanned aerial systems can be used to measure structural and phytochemical predictors of habitat quality at several spatial scales. We also demonstrate how advanced biotelemetry systems and robotic animals can be used to measure how habitat quality influences fine-scale habitat use, movement and reproductive effort of grouse. Integrating these technologies will allow researchers to better assess and manage the links among habitat quality (safety and food), resource acquisition (foraging behavior) and reproductive behaviors of grouse

Herpes simplex virus type 1 (HSV-1)-derived amplicon vectors for gene transfer and gene therapy

Amplicons are defective, helper -dependent, herpes simplex virus type 1 (HSV-1 )-derived vectors. The main interest of these vectors as gene transfer tools stems from the fact that the amplicon vector genomes do not carry protein-encoding viral sequences. Consequently, they are completely safe for the host and non-toxic for the infected cells. Moreover, the complete absence of virus genes provides space to accommodate very large foreign DNA sequences, up to almost 150-kbp, the size of the virus genome . This large transgene capacity can be used to deliver complete gene loci, including introns and exons, as well as long regulatory sequences, conferring tissue-specific expression, or stable maintenance of the transgene in proliferating cells. During many years the development of these vectors and their application in gene transfer experiments was hindered by the presence of contaminating toxic helper virus particles in the vector stocks. In recent years however, two different methodologies have been developed that allow generating amplicon stocks either completely free of helper particles or only faintly contaminated with fully defective helper particles. This chapter summarizes these two methodologies

The infectious BAC genomic DNA expression library: a high capacity vector system for functional genomics

Gene dosage plays a critical role in a range of cellular phenotypes, yet most cellular expression systems use heterologous cDNA-based vectors which express proteins well above physiological levels. In contrast, genomic DNA expression vectors generate physiologically-relevant levels of gene expression by carrying the whole genomic DNA locus of a gene including its regulatory elements. Here we describe the first genomic DNA expression library generated using the high-capacity herpes simplex virus-1 amplicon technology to deliver bacterial artificial chromosomes (BACs) into cells by viral transduction. The infectious BAC (iBAC) library contains 184,320 clones with an average insert size of 134.5 kb. We show in a Chinese hamster ovary (CHO) disease model cell line and mouse embryonic stem (ES) cells that this library can be used for genetic rescue studies in a range of contexts including the physiological restoration of Ldlr deficiency, and viral receptor expression. The iBAC library represents an important new genetic analysis tool openly available to the research community