JoVE Monthly Highlights: March 2018

1JoVE Content Production

Pooled shRNA Screen for Reactivation of MeCP2 on the Inactive X Chromosome

Vid Leko1,2, Smitha Sripathy1, Robin L. Adrianse1, Taylor Loe1, Angela Park1, Uyen Lao1, Eric J. Foss1, Marisa S. Bartolomei3, Antonio Bedalov1,4

We report a small hairpin RNA (shRNA) and next generation sequencing-based protocol for identifying regulators of X-chromosome inactivation in a murine cell line with firefly luciferase and hygromycin resistance genes fused to the methyl CpG binding protein 2 (MeCP2) gene on the inactive X chromosome.

A Method for Targeted 16S Sequencing of Human Milk Samples

Nicole H. Tobin1, Cora Woodward1, Sara Zabih1, David J. Lee1, Fan Li1, Grace M. Aldrovandi1

A semi-automated workflow is presented for targeted sequencing of 16S rRNA from human milk and other low-biomass sample types.

Practical Considerations in Studying Metastatic Lung Colonization in Osteosarcoma Using the Pulmonary Metastasis Assay

Michael M. Lizardo1,2, Poul H. Sorensen2,3

The goal of this article is to provide a detailed description of the protocol for the pulmonary metastasis assay (PuMA). This model permits researchers to study metastatic osteosarcoma (OS) cell growth in lung tissue using a widefield fluorescence or confocal laser-scanning microscope.

A Simple Method for High Throughput Chemical Screening in Caenorhabditis Elegans

Mark Lucanic1, Theo Garrett1, Matthew S. Gill2, Gordon J. Lithgow1

Here we describe a simple protocol for rapidly producing hundreds of nematode growth media agar, 96-well culture plates with consistent numbers of Caenorhhabditis elegans per well. These cultures are useful for the phenotypic screening of whole organisms. We focus here on using these cultures to screen chemicals for pro-longevity effects.