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A549 cells are adenocarcinomic human alveolar basal epithelial cells, and constitute a cell line that was first developed in 1972 by D. J. Giard, et al. through the removal and culturing of cancerous lung tissue in the explanted tumor of a 58-year-old caucasian male. [1]
The mutations required for immortality can occur naturally or be intentionally induced for experimental purposes. Immortal cell lines are a very important tool for research into the biochemistry and cell biology of multicellular organisms. Immortalised cell lines have also found uses in biotechnology.
Isogenic cell lines are created via a process called homologous gene-targeting. Targeting vectors that utilize homologous recombination are the tools or techniques that are used to knock-in or knock-out the desired disease-causing mutation or SNP (single nucleotide polymorphism) to be studied. Although disease mutations can be harvested ...
Types of mutations that can be introduced by random, site-directed, combinatorial, or insertional mutagenesis. In molecular biology, mutagenesis is an important laboratory technique whereby DNA mutations are deliberately engineered to produce libraries of mutant genes, proteins, strains of bacteria, or other genetically modified organisms. The ...
A549 cells share the same characteristics with the alveolar type II cells. This are type II pneumocytes crucial for lung homeostasis, and regeneration upon damage. For research purposes to unravel different molecular mechanisms leading to lung diseases, A549 cells serves as a good model to which translational research can be relied upon.
The mutation may be a single base change (a point mutation), multiple base changes, deletion, or insertion. The single-strand primer is then extended using a DNA polymerase, which copies the rest of the gene. The gene thus copied contains the mutated site, and is then introduced into a host cell in a vector and cloned.
Mutational robustness (also called mutation tolerance) describes the extent to which an organism's phenotype remains constant in spite of mutation. [9] Robustness can be empirically measured for several genomes [10] [11] and individual genes [12] by inducing mutations and measuring what proportion of mutants retain the same phenotype, function or fitness.
HRM offers a faster and more convenient closed-tube method of assessing the presence of mutations and gives a result which can be further investigated if it is of interest. In a study carried out by Scott et al. in 2006, [4] 3 cell lines harbouring different BRCA mutations were used to assess the HRM methodology. It was found that the melting ...