253,064 research outputs found
CHO microRNA engineering is growing up : recent successes and future challenges
No full textmicroRNAs with their ability to regulate complex pathways that control cellular behavior and phenotype have been proposed as potential targets for cell engineering in the context of optimization of biopharmaceutical production cell lines, specifically of Chinese Hamster Ovary cells. However, until recently, research was limited by a lack of genomic sequence information on this industrially important cell line. With the publication of the genomic sequence and other relevant data sets for CHO cells since 2011, the doors have been opened for an improved understanding of CHO cell physiology and for the development of the necessary tools for novel engineering strategies. In the present review we discuss both knowledge on the regulatory mechanisms of microRNAs obtained from other biological models and proof of concepts already performed on CHO cells, thus providing an outlook of potential applications of microRNA engineering in production cell lines
CHO-K1 B2M assay performance on CHO-K1 genomic DNA.
No full textThe CHO-K1 B2M qPCR assay was tested against a fourfold serial dilution of CHO-K1 genomic DNA spanning 0.025–102.4 ng gDNA. Graphing the Ct versus the log of plasmid copies demonstrates a linear range of 0.025–102.4 ng gDNA with R2 values approaching 1 and slope values close to -3.3.</p
CHO-S master cell line generation.
No full text(A) The H11 locus was cleaved, using CRISPR/Cas9, to encourage integration of the landing pad donor. A successful knock-in at the H11 locus generates a master cell line that has two PhiC31 attP sites, and that co-expresses three proteins via a PGK promoter-driven transcript. (B) Genotyping of the 5’-arm and 3’-arm in the CHO-S master cell line. PCR was performed with genomic DNA and specific primer pairs. 1: CHO-S parental cell line with 5’-arm primers; 2: CHO-S master cell line ("4–6") with 5’-arm primers; 3: CHO-S parental cell line with 3’-arm primers; 4: CHO-S master cell line ("4–6") with 3’-arm primers.</p
Structural and functional definition of the specificity of a novel caspase-3 inhibitor, Ac-DNLD-CHO-8
No full text<p><b>Copyright information:</b></p><p>Taken from "Structural and functional definition of the specificity of a novel caspase-3 inhibitor, Ac-DNLD-CHO"</p><p>http://www.biomedcentral.com/1471-2210/7/8</p><p>BMC Pharmacology 2007;7():8-8.</p><p>Published online 27 Jun 2007</p><p>PMCID:PMC1931592.</p><p></p>or 10 min with indicated concentrations of Ac-DNLD-CHO (●), Ac-DEVD-CHO (○), Ac-DQTD-CHO (△), or Ac-DMQD-CHO (□), and then the activities of the caspases were measured with each substrate as described in ''Methods''. The kinetics data presented are the means of three independent experiments
Structural and functional definition of the specificity of a novel caspase-3 inhibitor, Ac-DNLD-CHO-0
No full text<p><b>Copyright information:</b></p><p>Taken from "Structural and functional definition of the specificity of a novel caspase-3 inhibitor, Ac-DNLD-CHO"</p><p>http://www.biomedcentral.com/1471-2210/7/8</p><p>BMC Pharmacology 2007;7():8-8.</p><p>Published online 27 Jun 2007</p><p>PMCID:PMC1931592.</p><p></p>or 10 min with indicated concentrations of Ac-DNLD-CHO (●), Ac-DEVD-CHO (○), Ac-DQTD-CHO (△), or Ac-DMQD-CHO (□), and then the activities of the caspases were measured with each substrate as described in ''Methods''. The kinetics data presented are the means of three independent experiments
Unimodality of Betti numbers for Hamiltonian circle actions with index-increasing moment Maps
No full textThe unimodality conjecture posed by Tolman in [L. Jeffrey, T. Holm, Y. Karshon, E. Lerman and E. Meinrenken, Moment maps in various geometries, http://www.birs.ca/workshops/2005/05w5072/report05w5072.pdf] states that if (M,ω) is a 2n-dimensional smooth compact symplectic manifold equipped with a Hamiltonian circle action with only isolated fixed points, then the sequence of Betti numbers {b0(M),b2(M),...,b2n(M)} is unimodal, i.e. bi(M) ≤ bi+2(M) for every i < n. Recently, the author and Kim [Y. Cho and M. Kim, Unimodality of the Betti numbers for Hamiltonian circle action with isolated fixed points, Math. Res. Lett. 21(4) (2014) 691-696] proved that the unimodality holds in eight-dimensional case by using equivariant cohomology theory. In this paper, we generalize the idea in [Y. Cho and M. Kim, Unimodality of the Betti numbers for Hamiltonian circle action with isolated fixed points, Math. Res. Lett. 21(4) (2014) 691-696] to an arbitrary dimensional case. We prove the conjecture in arbitrary dimension under the assumption that the moment map H : M → R is index-increasing, which means that ind(p) < ind(q) implies H(p) < H(q) for every pair of critical points p and q of H, where ind(p) is the Morse index of p with respect to H. © World Scientific Publishing Company1111sciescopu
Cho et al. 2022 DoLP observations
No full textDataset (PM2.5-DoLP relationship) for Cho et al. 2022</p
CHO-K1, COS7, and HEK293 cells express PAR1 and PAR2 receptors.
No full textA. CHO-K1, COS7, and HEK293 cells naturally express high levels of PAR1 and PAR2 mRNA but express little or no PAR3 and PAR4 mRNA. qPCR analysis was used to quantify the mRNA expression. Specific primers for each of PAR1, PAR2, PAR3, and PAR4, were used to quantify the respective mRNA expression using cDNA made from each cell line as the templates. β-actin primers were used to quantify β-actin mRNA expression as the internal controls. The relative mRNA expression of PAR1, PAR2, PAR3, and PAR4 were first normalized using β-actin expression, and then normalized using the PAR1 expression level in CHO-K1 cells, which is arbitrarily set as 100%. The relative expressions of other genes are represented as percentage of PAR1 mRNA level in CHO-K1 cells. The results shown are mean ± sd (n = 3). Statistical analysis (One-Way ANOVA) shows that compared with the mRNA expression of PAR4, which is undetectable in these cells, CHO cells express high levels of mRNAs for PAR1 (** p = 0.0037), PAR2 (* p = 0.023), and PAR3 (* p = 0.035); COS7 and HEK293 cells express high level of mRNAs for PAR1 (** p = 0.0029, * p = 0.032, respectively) and PAR2 (** p = 0.0013, ** p = 0.0027, respectively) without expressing detectable PAR3 and PAR4 mRNAs. B, C, D. CHO-K1, COS7, and HEK293 cells naturally express PAR1 and PAR2 receptors and respond to thrombin (PAR1 ligand) and trypsin (PAR2 ligand) stimulations. FLIPR assays were used measure receptor activation as indicated by intracellular Ca2+ mobilization. Relative fluorescent units (RFU) are the readout for fluorescent intensities for Ca2+ mobilization signals. Various concentration of thrombin or trypsin were used as the ligands to activation the receptors. The assays were performed in triplicates at each data point and mean ± sd are shown. E. Sequencing analysis of the genomic DNA from par1 & par2 knock out HEK293 cells. The results show that a 270 bp deletion in par1 gene and a 347 bp deletion in par2 gene have been achieved. The deletions removed the coding regions from TM2 to TM3 for both PAR1 and PAR2 proteins. The vertical lines indicate the deletion sites. F. Characterization of par1 & par2 knock-out HEK293 cells. FLIPR assays were used to characterize receptor activation as indicated. Wild type HEK293 cells were used as the positive control. The assays were performed in triplicates at each data point and mean ± sd are shown.</p
Structural and functional definition of the specificity of a novel caspase-3 inhibitor, Ac-DNLD-CHO-1
No full text<p><b>Copyright information:</b></p><p>Taken from "Structural and functional definition of the specificity of a novel caspase-3 inhibitor, Ac-DNLD-CHO"</p><p>http://www.biomedcentral.com/1471-2210/7/8</p><p>BMC Pharmacology 2007;7():8-8.</p><p>Published online 27 Jun 2007</p><p>PMCID:PMC1931592.</p><p></p>etric caspase substrates Ac-DNLD-MCA (●) and Ac-DEVD-CHO (○) were compared. The cleavage assay was performed as described in "Methods" The y-axis represents the concentration of MCA production (pmol) and the x-axis represents incubation period. Data indicate the mean of three independent experiments
Structural and functional definition of the specificity of a novel caspase-3 inhibitor, Ac-DNLD-CHO-6
No full text<p><b>Copyright information:</b></p><p>Taken from "Structural and functional definition of the specificity of a novel caspase-3 inhibitor, Ac-DNLD-CHO"</p><p>http://www.biomedcentral.com/1471-2210/7/8</p><p>BMC Pharmacology 2007;7():8-8.</p><p>Published online 27 Jun 2007</p><p>PMCID:PMC1931592.</p><p></p>tively. Hydrogen bonds are shown as dashed lines. Hydrophobic interactions are shown as thick broken lines schematically. A, The binding mode of Ac-DNLD-CHO was obtained from docking simulations. B, The binding mode of Ac-DEVD-CHO was obtained from the X-ray crystal structure (1PAU). C, The time courses of liberation of fluorescence (MCA) from Ac-DNLD-MCA catalyzed by wild-type caspase-3 (●) and substituted (S209A) caspase-3 (▲). D, The time courses of liberation of fluorescence (MCA) from Ac-DEVD-MCA catalyzed by wild-type caspase-3 (○) and substituted (S209A) caspase-3 (△). The cleavage assays were performed as described in ''Methods''. Data indicate the mean of three independent experiments. E, Amounts of wild-type (lane 2) and substituted (S209A) (lane 3) active caspase-3 proteins generated by coexpression of HA-p17 and HA-p12 subunits in in vitro translation system were analyzed by Western blotting as described under ''Methods''. In this experiment, empty vector was used as control (lane 1)
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