2 research outputs found
Biomarkers to identify and isolate senescent cells.
Aging is the main risk factor for many degenerative diseases and declining health. Senescent cells are part of the underlying mechanism for time-dependent tissue dysfunction. These cells can negatively affect neighbouring cells through an altered secretory phenotype: the senescence-associated secretory phenotype (SASP). The SASP induces senescence in healthy cells, promotes tumour formation and progression, and contributes to other age-related diseases such as atherosclerosis, immune-senescence and neurodegeneration. Removal of senescent cells was recently demonstrated to delay age-related degeneration and extend lifespan. To better understand cell aging and to reap the benefits of senescent cell removal, it is necessary to have a reliable biomarker to identify these cells. Following an introduction to cellular senescence, we discuss several classes of biomarkers in the context of their utility in identifying and/or removing senescent cells from tissues. Although senescence can be induced by a variety of stimuli, senescent cells share some characteristics that enable their identification both in vitro and in vivo. Nevertheless, it may prove difficult to identify a single biomarker capable of distinguishing senescence in all cell types. Therefore, this will not be a comprehensive review of all senescence biomarkers but rather an outlook on technologies and markers that are most suitable to identify and isolate senescent cells
EMMA: An Extensible Mammalian Modular Assembly Toolkit for the Rapid Design and Production of Diverse Expression Vectors
Mammalian
plasmid expression vectors are critical reagents underpinning
many facets of research across biology, biomedical research, and the
biotechnology industry. Traditional cloning methods often require
laborious manual design and assembly of plasmids using tailored sequential
cloning steps. This process can be protracted, complicated, expensive,
and error-prone. New tools and strategies that facilitate the efficient
design and production of bespoke vectors would help relieve a current
bottleneck for researchers. To address this, we have developed an
extensible mammalian modular assembly kit (EMMA). This enables rapid
and efficient modular assembly of mammalian expression vectors in
a one-tube, one-step golden-gate cloning reaction, using a standardized
library of compatible genetic parts. The high modularity, flexibility,
and extensibility of EMMA provide a simple method for the production
of functionally diverse mammalian expression vectors. We demonstrate
the value of this toolkit by constructing and validating a range of
representative vectors, such as transient and stable expression vectors
(transposon based vectors), targeting vectors, inducible systems,
polycistronic expression cassettes, fusion proteins, and fluorescent
reporters. The method also supports simple assembly combinatorial
libraries and hierarchical assembly for production of larger multigenetic
cargos. In summary, EMMA is compatible with automated production,
and novel genetic parts can be easily incorporated, providing new
opportunities for mammalian synthetic biology