细胞技术

The in vitro translation system provides an important means of identifying mRNA species of a gene of interest, characterizing the protein products, and investigating translational control. Rabbit reticulocyte lysate (RRL) or wheat germ lysate have been used to successfully transl ...

The procedure described here, the PCR poly(A) test (PAT), allows the analysis of the poly(A) tail on a specific mRNA within a pool of total RNA in a rapid (1 day) and sensitive (subnanogram total RNA) fashion. The assay also provides quantitative estimates of the RNAs poly(A) tail length (1). RNA manipulatio ...

The 3′ ends of most nonhistone mRNAs in mammalian cells are generated by the endonucleolytic cleavage of an mRNA precursor (pre-mRNA) followed by the addition of a polyadenylate (poly) tail (see ref. 1 for a recent review on mRNA 3′ end processing). The ability to process pre-mRNAs in vitro has contribu ...

In recent years, the SR protein family of precursor messenger RNA splicing factors has emerged as a key player in the assembly of the spliceosomal machinery onto pre-mRNA. The SR proteins are essential splicing factors and different family members can direct usage of alternative splice sites ...

Pluripotent embryonic stem (ES) cell lines were first isolated over 25 years ago and remain an essential tool in molecular and developmental biology to this day. In particular, the use of homologous recombination and subsequent generation of ES-derived mice has greatly facilitated rese ...

Human embryonic stem cells (hESCs) represent a powerful platform to study human development and its dysfunction in human disease. However, certain biological properties have hampered the application of standard gain of function and loss of function tools to these cells. For example, wh ...

Local micro-environmental cues consisting of soluble cytokines, extra-cellular matrix (ECM), and cell–cell contacts are determining factors in stem cell fate. These extrinsic cues form a ‘niche’ that governs a stem cell’s decision to either self-renew or differentiate into one or more c ...

The ability of cell-penetrating peptides (CPPs) to cross cell membranes and transport cargo into cells makes them an attractive tool for the molecular engineering of stem cells. Even though the exact mechanism of transduction remains unclear, their potential has been demonstrated for d ...

Tissue-resident stem cells or primitive progenitors play an integral role in homeostasis of most organ systems. Recent developments in methodologies to isolate and culture embryonic and somatic stem cells have many new applications poised for clinical and preclinical trials, whi ...

The identification and cloning of thrombopoietin was certainly a defining moment for the study of megakaryopoiesis and thrombopoiesis ex vivo. This and other progresses made in the development of culture processes for hematopoietic stem cells have paved the way for ongoing clinical t ...

Interactions between haemopoietic stem cells (HSC) and their microenvironment serve multiple functions including the attraction to and retention and regulation in the bone marrow HSC niche. However, the cell adhesion molecules involved, their HSC receptors and the mechanisms un ...

A growing number of specific cell types have been successfully derived from embryonic stem cells (ES cells), including a variety of neural cells. In vitro generated cells need to be extensively characterized to establish functional equivalency with their in vivo counterparts. The ultim ...

We describe a technological approach permitting the massive expansion of CD34+ stem cells and their 100% conversion ex vivo into mature red blood cells (RBC). The protocol comprises three steps: a first step consisting of cell proliferation and induction of erythroid differentiation in a l ...

The isolation and experimental manipulation of multipotent precursors is of increasing therapeutic relevance. We recently reported the generation of cultures of Skin-derived Precursors (‘SKPs’), multipotent cells that can be isolated from the dermis of embryonic, neonatal, and ...

Neural transplantation has been a long-standing goal for the treatment of neurological injury and disease. The recent discovery of persistent pools of neural stem cells within the adult mammalian brain has reignited interest in transplant therapeutics. Since neural stem cells are se ...

Islet grafts have demonstrated that patients with diabetes would benefit greatly by β-cell therapy. However, the paucity of available islets for transplantation as well as the immunological barriers faced in allogeneic transplantation represent a tremendous barrier to regene ...

Stem cells are present in all tissues and organs, and are crucial for normal regulated growth. How the pool size of stem cells and their progeny is regulated to establish the tissue prenatally, then maintain it throughout life, is a key question in biology and medicine. The ability to precisely locate s ...

Researchers interested in studying RNA structure and function or RNA-protein interactions are increasingly using site-specifically modified RNAs to probe sites of interest (1). In addition to expanding the repertoire of functional groups beyond the very limited set available th ...

A large number of genes express multiple mRNAs that encode diverse protein isoforms via alternative pre-mRNA splicing. For many genes, alternative splicing is regulated according to cell-specific patterns (in this chapter, cell-specific is used as a general term to refer to regulation a ...

The spliceosome is a large ribonucleoprotein that splices precursor messenger RNA (pre-mRNA) in the nuclei of eukaryotic cells (reviewed in refs. 1–3). It is composed of five small nuclear ribonucleoproteins (snRNPs) and numerous non-snRNP proteins. Each snRNP consists of a small nucle ...