细胞功能测定

DNA mismatch repair plays an important role in mutation avoidance by recognizing and correcting mismatched bases and loops prior to their fixation as mutations. When mismatch repair is defective, cells exhibit elevated rates of spontaneous mutations. For example, microsatellite s ...

During the last 20 years, the cloning and identification of DNA repair genes in bacteria, yeast, human, mouse, and other organisms have been an expanding enterprise. Gene/cDNA isolation and analysis is a critical step toward discovering gene/protein function by enabling subsequent char ...

Cell lines with an increased sensitivity to mutagens, such as ultraviolet (UV) light, X-rays, alkylating compounds, and crosslinking agents, are defective in a cellular response to these agents. These responses include mechanisms that process DNA lesions, scavenge free radicals, or re ...

This chapter describes a method to isolate γ-ray hypersensitive mutants of Arabidopsis thaliana from ethyl methanesulfonate- (EMS) treated seed (1). The mutants are identified by visible symptoms of extreme radiation damage 10–15 d following exposure of seedlings to a threshold 10-k ...

UV radiation induces two major DNA damage products: the cyclobutane pyrimidine dimer (CPD) and the pyrimidinepyrimidinone dimer (or photoproduct; PP). The biological effects of both lesions have been studied in microbial and mammalian systems. Pyrimidine dimers have been shown to act ...

The fruitfly Drosophila melanogaster offers numerous advantages as a metazoan model for genetic dissection of conserved biological processes, such as DNA repair. Its ease of culture, short generation time, small number of linkage groups (2n=8), and giant polytene chromosomes, combi ...

The nematode Caenorhabditis elegans has gained widespread popularity for use in addressing many biological problems, particularly those relating to development (for brief topical reviews, see 1–5; for comprehensive treatises, see 6–10). This can be attributed to both inherent pro ...

Eukaryotic cells have the ability to influence progression through the cell cycle in response to internal and external inputs of “information”. They do so by using feedback control mechanisms able to arrest mitosis in response to different cellular events. Such active mechanisms capab ...

Considerations in rational designs of CPP-based transcutaneous delivery systems are described. Impact of design considerations of nonclinical and clinical results are presented in detail.

Since the discovery over 15 years ago of a protein transcription factor that possessed the ability to cross the plasma membrane, cell-penetrating peptides (CPPs) have been evaluated for the ability to transport diverse cargoes into cells, tissues, and organs. Certain CPPs have been used for t ...

To enhance the cytosolic delivery of therapeutic drugs and genes, pH-sensitive and membrane fusiogenic peptides have been employed as additives for facilitating their endosomal escape. GALA is such a peptide composed of repeating sequences of Glu-Ala-Leu-Ala, which are designed to m ...

This chapter describes the manufacturing process to a certain level for a possible oligonucleotide cargo and a peptide API in a multi-kilogram scale from a manufacture’s point of view. In the concluding remarks, possible conjugation methods will be discussed from an industrial-scale pe ...

A major challenge confronting the further advancement of using molecular transporters conjugated to small molecular weight therapeutics in the clinic is the development of linkers that would allow for the controllable release of a free drug/probe only after cell entry. Development of ...

RNA-binding proteins (RBPs) are fundamental regulatory proteins for all forms of transcriptional and posttranscriptional control of gene expression. However, isolating RBPs is technically challenging for investigators. Currently, the most widely used techniques to isol ...

The first series of cell-penetrating penta-peptides (CPP5s) were discovered as cytoprotective penta-peptides designed from the Bax-inhibiting domain of Ku70. Bax is an inducer of programmed cell death, and Ku70 is a multifunctional protein maintaining genomic stability and pro ...

Cell-penetrating peptides (CPPs) are a very interesting class of molecules to be introduced in gene and siRNA vectors. They can be used to overcome one of the biggest hurdles in gene and siRNA delivery in vitro and in vivo, the transfer across cell membranes. This chapter describes protocols for the s ...

Cell-penetrating peptides (CPPs), in particular TATp, have been widely used for intracellular delivery of various cargoes, both in vitro and in vivo. Modifications of nanoparticles with CPPs require either covalent or noncovalent approach. Here we describe various methods to attach ...

Two of the most promising and complex areas in biologics development, either as research tools or potential therapeutics, are cell-penetrating peptides (CPPs) and RNA interference (RNAi) modulators. Consequently, the combined application of these technologies in pursuit of imp ...

Each normal organ and pathological condition expresses a distinct set of molecules on their vasculature. These molecular signatures have been efficiently profiled using in vivo phage display technology. Using this technology, several peptides homing specifically to tumour bl ...

Cell-penetrating peptides (CPPs) have been widely used for a cellular delivery of biologically relevant cargoes including antisense peptide nucleic acids (PNAs). Although chemical conjugation of PNA to a variety of CPPs significantly improves the cellular uptake of the PNAs, bioa ...