01-05-2013 | Joseph T.P. Yeeles; Jérôme Poli; Kenneth J. Marians; Philippe Pasero, CSH Perspectives, 2013

In recent years, an increasing number of studies have shown that prokaryotes and eukaryotes are armed with sophisticated mechanisms to restart stalled or collapsed replication forks. Although these processes are better understood in bacteria, major breakthroughs have also been made to ...

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01-05-2013 | Verena Pfeiffer; Joachim Lingner, CSH Perspectives, 2013

Telomeres are the physical ends of eukaryotic chromosomes. They protect chromosome ends from DNA degradation, recombination, and DNA end fusions, and they are important for nuclear architecture. Telomeres provide a mechanism for their replication by semiconservative DNA replication and ...

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01-05-2013 | Lynette C. Foo, CSH Protocols, 2013

We describe the use of immunopanning to purify rodent astrocytes. Immunopanning of astrocytes permits the prospective isolation of astrocytes from the rodent brain. Prospective isolation refers to the direct selection of cells without multiple steps that extend over a few days, thereby ...

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01-05-2013 | Mikhail Alexeyev; Inna Shokolenko; Glenn Wilson; Susan LeDoux, CSH Perspectives, 2013

DNA molecules in mitochondria, just like those in the nucleus of eukaryotic cells, are constantly damaged by noxious agents. Eukaryotic cells have developed efficient mechanisms to deal with this assault. The process of DNA repair in mitochondria, initially believed nonexistent, has now ...

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01-05-2013 | Kishore K. Chiruvella; Zhuobin Liang; Thomas E. Wilson, CSH Perspectives, 2013

Nonhomologous end joining (NHEJ) refers to a set of genome maintenance pathways in which two DNA double-strand break (DSB) ends are (re)joined by apposition, processing, and ligation without the use of extended homology to guide repair. Canonical NHEJ (c-NHEJ) is a well-defined pathway with ...

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01-05-2013 | Jain, R. K., Munn, L. L., Fukumura, D., CSH Protocols, 2013

Noninvasive techniques have been developed for the assessment of vascular parameters, including vascular permeability, in normal and diseased tissues of mice. In this protocol, mice are injected with an appropriate fluorescent tracer. The effective average vascular permeability (P) of a ...

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01-05-2013 | Henning Walczak, CSH Perspectives, 2013

The discovery of tumor necrosis factor (TNF) marked the beginning of one of the most fascinating journeys in modern biomedical research. For the moment, this journey has culminated in the development of drugs that inhibit TNF. TNF blockers have revolutionized the treatment of many chronic ...

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01-05-2013 | Carey, M. F., Peterson, C. L., Smale, S. T., CSH Protocols, 2013

DNase I footprinting has found a wide following for both identifying and characterizing DNA–protein interactions, particularly because of its simplicity. The concept is that a partial digestion by DNase I of a uniquely 32P-end-labeled fragment will generate a ladder of fragments, whose ...

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01-05-2013 | Steven J. Burden; Norihiro Yumoto; Wei Zhang, CSH Perspectives, 2013

Muscle-specific kinase (MuSK) is essential for each step in neuromuscular synapse formation. Before innervation, MuSK initiates postsynaptic differentiation, priming the muscle for synapse formation. Approaching motor axons recognize the primed, or prepatterned, region of muscle, causing ...

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01-05-2013 | Zhou, R., Mohr, S., Hannon, G. J., Perrimon, N., CSH Protocols, 2013

In Drosophila cells, RNA interference (RNAi) can be triggered by synthetic long double-stranded RNAs (dsRNAs). For many Drosophila cell lines and cell types, passive dsRNA uptake is inefficient. More complete silencing responses can often be obtained in Drosophila S2 cells using ...

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