细胞功能测定

Gene therapy is a promising approach for the treatment of a variety of disorders including genetic diseases and cancer. Among the viral vectors used in gene therapy, the lentiviral vector, based on HIV-1, is the only integrative vector able to transduce nondividing cells. The first generation of ...

Gene delivery vectors based on recombinant adeno-associated virus (AAV) are powerful tools for studying myogenesis in normal and diseased conditions. Strategies have been developed to use AAV to increase, down-regulate, or modify expression of a particular muscle gene in a specific mu ...

β-galactosidase (β-gal) is among the most frequently used markers for studying a wide variety of biological mechanisms, e.g., gene expression, cell migration, stem cell conversion to different cell types, and gene silencing. Many of these studies require the histochemical detection of r ...

The use of nonviral (plasmid DNA) gene delivery into skeletal muscle has increased significantly in recent years. The procedure is used to overexpress wild-type proteins, express mutant proteins, or knock down endogenous proteins. These manipulations can identify the role of a specific ...

MicroRNAs (miRNAs) are a class of small ∼22 nucleotide noncoding RNAs which regulate gene expression at the posttranscriptional level by either destabilizing and consequently degrading their targeted mRNAs or by repressing their translation. Emerging evidence has demonstra ...

RNA interference appears as a promising tool for therapeutic gene silencing to block protein expression. A long-lived silencing is obtained through the in situ expression of shRNA. A safe approach is to use a physical method such as in vivo electropulsation with plate electrodes. This is prese ...

An electrophoretic mobility shift assay (EMSA) is a common and invaluable technique which can be utilized to study the affinity of proteins to a specific DNA or RNA sequence. These assays are performed in vitro with protein extracts isolated from either cultured cells or isolated tissues. Here, ...

Secondary skeletal muscle fiber phenotype is dependent upon depolarization from motor neuron innervation. To study the effects of depolarization on muscle fiber type development, several in vivo and in vitro model systems exist. We have developed a relatively simple-to-use in vitro m ...

Mesoangioblasts (MABs) are mesoderm-derived stem cells, associated with small vessels and originally described in the mouse embryonic dorsal aorta. Similar though not identical cells have been later identified and characterized from postnatal small vessels of skeletal muscle ...

Satellite cells are a heterogeneous population of muscle progenitors with stem cell properties responsible for the regeneration of adult skeletal muscle. Increasing interest in the therapeutic potential of satellite cells has challenged researchers with the need to purify a hom ...

Repair of adult skeletal muscle depends on satellite cells, myogenic stem cells located between the basal lamina and the plasmalemma of the myofiber. Standardized protocols for the isolation and culture of satellite cells are key tools for understanding cell autonomous and extrinsic f ...

Dissociated human fetal skeletal muscle contains myogenic cells, as well as non-myogenic cells such as adipocytes, fibroblasts, and lymphocytes. It is therefore important to determine an efficient and reliable isolation method to obtain a purer population of myoblasts. Toward this e ...

Mapping the cis-regulatory modules (CRMs) to which bind myogenic transcription factors is an �obligatory step towards understanding gene regulatory networks governing muscle development and function. This can be achieved in silico or by chromatin immunoprecipitation (ChIP) ...

Studies investigating mechanisms that control gene regulation frequently examine the accessibility of specific DNA sequences to nuclease cleavage. In general, sequences that are sensitive to nuclease cleavage are considered to be in an “open” chromatin conformation that is ass ...

Studies investigating mechanisms controlling gene regulation frequently examine specific DNA sequences using chromatin immunoprecipitation (ChIP) assays to determine whether specific regulatory factors or modified histones are present. While use of primary cells or c ...

In order to determine how gene expression is regulated in response to environmental cues, it is necessary to identify the specific interaction between transcription factors and their cognate cis-regulatory DNA elements. Here we have out-lined electrophoretic mobility shift assay ...

Detection of transcription factors in immune cell populations, particularly in subpopulations that are represented at low frequencies in lymphoid and nonlymphoid organs, presents a particular challenge when using traditional methods such as western blot analysis. Therefo ...

In the age of systems biology, biologists seek to quantify the absolute number of molecules in experimentally treated samples. Immunoblotting remains a technique of choice for assessing the relative differences between the protein levels in different samples. Here we discuss how to ex ...

In response to environmental stress, cells trigger a number of molecular mechanisms to control their survival and growth. These include changes in gene expression with corresponding Post-translational modifications to critical transcriptional-control proteins. Trans ...

The chromatin organizer SATB1 regulates distant genes by selectively tethering matrix attachment regions (MARs) to the nuclear matrix. Post-translational modifications (PTMs) are important regulators of functional activities of proteins. Recently, a phosphorylation ...