Assessment of Cell Viability

Johnson, Simon C.; Nguyen, Vy; Coder, David M.

doi:10.1002/0471142956.cy0902s64

Cited by 89 publications

(63 citation statements)

References 59 publications

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“…Cells were resuspended in 1 mL of HSM with 5 mM of the vital fluorescent dye carboxymethylfluorescein diacetate (CMFDA; Invitrogen; Johnson et al, 2013). Cells were kept in the dark at room temperature for 30 to 40 min and then washed two times with phosphate-buffered saline (PBS).…”

Section: Cell Viability Analysismentioning

confidence: 99%

Defects in a New Class of Sulfate/Anion Transporter Link Sulfur Acclimation Responses to Intracellular Glutathione Levels and Cell Cycle Control

et al. 2014

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We previously identified a mutation, suppressor of mating type locus3 15-1 (smt15-1), that partially suppresses the cell cycle defects caused by loss of the retinoblastoma tumor suppressor-related protein encoded by the MAT3 gene in Chlamydomonas reinhardtii. smt15-1 single mutants were also found to have a cell cycle defect leading to a small-cell phenotype. SMT15 belongs to a previously uncharacterized subfamily of putative membrane-localized sulfate/anion transporters that contain a sulfate transporter domain and are found in a widely distributed subset of eukaryotes and bacteria. Although we observed that smt15-1 has a defect in acclimation to sulfur-limited growth conditions, sulfur acclimation (sac) mutants, which are more severely defective for acclimation to sulfur limitation, do not have cell cycle defects and cannot suppress mat3. Moreover, we found that smt15-1, but not sac mutants, overaccumulates glutathione. In wild-type cells, glutathione fluctuated during the cell cycle, with highest levels in mid G1 phase and lower levels during S and M phases, while in smt15-1, glutathione levels remained elevated during S and M. In addition to increased total glutathione levels, smt15-1 cells had an increased reduced-to-oxidized glutathione redox ratio throughout the cell cycle. These data suggest a role for SMT15 in maintaining glutathione homeostasis that impacts the cell cycle and sulfur acclimation responses.

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Section: Cell Viability Analysismentioning

confidence: 99%

Defects in a New Class of Sulfate/Anion Transporter Link Sulfur Acclimation Responses to Intracellular Glutathione Levels and Cell Cycle Control

et al. 2014

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“…Treatment of cells with a cytotoxic compound can stall cell growth and eventually lead to cell death, which can be determined by cell viability assays. [1][2][3] These assays have been extensively used in lead discovery for identification of active compounds for cancer and many infectious pathogens. Cell viability assays have also been used to assess compound cytotoxicity for toxicological studies.…”

Section: Introductionmentioning

confidence: 99%

High-Throughput Viability Assay Using an Autonomously Bioluminescent Cell Line with a Bacterial Lux Reporter

et al. 2015

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“…After puromycin selection, the cells in each well were divided into two groups. One group was used to calculate the survival rate using propidium iodide (PI) staining as previously described [24]. The other group was grown in fresh medium without puromycin for 2~3 days and genomic DNA was extracted for further mutation detection.…”

Section: Methodsmentioning

confidence: 99%

Enhancing Targeted Genomic DNA Editing in Chicken Cells Using the CRISPR/Cas9 System

Yang

Guo²,

et al. 2017

PLoS ONE

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The CRISPR/Cas9 system has enabled highly efficient genome targeted editing for various organisms. However, few studies have focused on CRISPR/Cas9 nuclease-mediated chicken genome editing compared with mammalian genomes. The current study combined CRISPR with yeast Rad52 (yRad52) to enhance targeted genomic DNA editing in chicken DF-1 cells. The efficiency of CRISPR/Cas9 nuclease-induced targeted mutations in the chicken genome was increased to 41.9% via the enrichment of the dual-reporter surrogate system. In addition, the combined effect of CRISPR nuclease and yRad52 dramatically increased the efficiency of the targeted substitution in the myostatin gene using 50-mer oligodeoxynucleotides (ssODN) as the donor DNA, resulting in a 36.7% editing efficiency after puromycin selection. Furthermore, based on the effect of yRad52, the frequency of exogenous gene integration in the chicken genome was more than 3-fold higher than that without yRad52. Collectively, these results suggest that ssODN is an ideal donor DNA for targeted substitution and that CRISPR/Cas9 combined with yRad52 significantly enhances chicken genome editing. These findings could be extensively applied in other organisms.

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Assessment of Cell Viability

Cited by 89 publications

References 59 publications

Defects in a New Class of Sulfate/Anion Transporter Link Sulfur Acclimation Responses to Intracellular Glutathione Levels and Cell Cycle Control

Defects in a New Class of Sulfate/Anion Transporter Link Sulfur Acclimation Responses to Intracellular Glutathione Levels and Cell Cycle Control

High-Throughput Viability Assay Using an Autonomously Bioluminescent Cell Line with a Bacterial Lux Reporter

Enhancing Targeted Genomic DNA Editing in Chicken Cells Using the CRISPR/Cas9 System

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