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Chao, W., Kolski-Andreaco, A. March 2016 - This Month in JoVE: RNA interference in Mosquitoes, iPSCs Derived from Nasal Epithelia, Air Sampling of Atmospheric Aerosols, and Autologous Micro-grafts for Skin Lesions. J. Vis. Exp. (109), e5772, (2016).
In JoVE Biology, RNA interference (RNAi) is a natural mechanism of gene silencing that occurs via double-stranded RNA (dsRNA), which targets homologous DNA for degradation. This phenomenon allows researchers to selectively silence genes in many eukaryotes-making RNAi an extremely valuable tool for uncovering gene function. However, in the mosquito Anopheles gambiae, a major vector for malaria, RNAi has limited ability to target genes during developmental stages. This month, Regna et al. present an RNAi protocol using direct injection during pupal development. After the pupae complete development, their adult phenotypes confirm the gene knockdown. This method expands the arsenal of genomic tools for vector insect research.
In JoVE Developmental Biology, induced pluripotent stem cells (iPSCs) have been generated from many cell types, and are valuable models for human development and disease. iPSCs are also valued for their potential applications in regenerative medicine. This month, Ulm et al. demonstrate methods for sampling nasal epithelial mucosa from children, then culturing the samples to obtain nasal epithelial cells (NECs), which are reprogrammed into iPSCs. NECs are of particular interest because they're the primary cells infected with respiratory viruses, and are readily accessible during clinical visits. Therefore, this protocol facilitates patient-specific research in airway epithelial biology.
In JoVE Environment, filters are important tools in atmospheric aerosols research. These sampled filters collect ambient particles, such as endotoxins and biological aerosols, for analysis. In this issue, Lang-Yona et al. use air-sampled filters for two complementary analyses of atmospheric biological particles: endotoxin and DNA. Specifically, they study endotoxin components of gram-negative bacterial cell walls, known collectively as lipopolysaccharide (LPS). In parallel, they perform a genomic evaluation of the sample's bacterial content. This method can produce highly accurate and reliable analyses for biological aerosol research.
In JoVE Medicine, researchers are finding new ways to treat wounds with a patient's own tissues, This month, Purpura et al. describe a new method of creating autologous micrografts. When cultured on collagen sponges, these micro-grafts become bio-complexes ready to use in the treatment of skin lesions. These biocomplexes were applied in a patient, who showed good healing after 30 days. This new regenerative approach shows promise as an efficient, one-step treatment of acute and chronic lesions.
Valeria Purpura1, Elena Bondioli1, Antonio Graziano2, Letizia Trovato2, Davide Melandri1, Martina Ghetti1, Andrea Marchesini3, Maria Gabriella Cusella De Angelis4,5, Laura Benedetti4,5, Gabriele Ceccarelli4,5, Michele Riccio3
1Burns Centre and Emilia Romagna Regional Skin Bank, 2Human Brain Wave srl, 3Plastic and Reconstructive Surgery, AOU “Ospedali Riuniti”, 4Department of Public Health, Experimental Medicine, Anatomy and Forensic, University of Pavia, 5C.H.T Centre for Health Technologies, University of Pavia
The protocol describes a new method to disaggregate human tissues and to create autologous micro-grafts that, combined with collagen sponges, give rise to human bio-complexes ready to use in the treatment of skin lesions. Further, this system preserves cell viability of micro-grafts at different times after mechanical disaggregation.
Naama Lang-Yona1,2, Yinon Mazar1, Michal Pardo1, Yinon Rudich1
1Department of Earth and Planetary Sciences, Weizmann Institute of Science, 2Multiphase Chemistry Department, Max Planck Institute
Two complementary analyses of atmospheric biological particles from air sampled filters are described herein: the extraction and detection of endotoxin, and of DNA.
Kimberly Regna1, Rachel M. Harrison1, Shannon A. Heyse1, Thomas C. Chiles1, Kristin Michel2, Marc A. T. Muskavitch1,3
1Biology Department, Boston College, 2Division of Biology, Kansas State University, 3Discovery Research, Biogen
RNA interference (RNAi) is an extremely valuable tool for uncovering gene function. However, the ability to target genes using RNAi during pre-adult stages is limited in the major human malaria vector Anopheles gambiae. We describe an RNAi protocol to reduce gene function via direct injection during pupal development.
Cultivate Primary Nasal Epithelial Cells from Children and Reprogram into Induced Pluripotent Stem Cells
Ashley Ulm1, Christopher N. Mayhew2, Jason Debley3, Gurjit K. Khurana Hershey4, Hong Ji1,4
1Pyrosequencing Core, Cincinnati Children's Hospital, 2Division of Developmental Biology, Cincinnati Children's Hospital, 3Division of Pulmonary Medicine, Seattle Children's Hospital, 4Division of Asthma Research, Cincinnati Children's Hospital
This publication demonstrates methods for successful sampling and culture of nasal epithelial mucosa from children, and reprogramming these cells to induced Pluripotent Stem Cells (iPSCs).
No conflicts of interest declared.