Global ETD Search
Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
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Showing 1 to 19 of 19 for “"DT40"”.
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Disruption of UHRF1 expression in chicken DT40 cells
… in the most effective gene targeting system of DT40 cells to observe the effect of UHRF1 deletion on DNA replication and the cellular response to DNA damage. CRISPR/Cas9 technology and traditional gene targeting were applied to generate conditional UHRF1 knockout cell lines. It was found that …
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Gene Targeting in a Novel Mouse Model and the Chicken DT40 Cell Line
… most cells; an exception to this is the chicken DT40 cell line which has a high relative rate of gene targeting (gene targeting rate/ random integration rate). We therefore sought to determine the basis of this high rate of gene targeting using assays which quantitate the rates of repairing DNA …
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In vitro diversification of chimeric human/chicken antibodies
… ones (Cumbers et al., 2002). The chicken derived DT40 B cell line has been adapted for the expression and diversification of humanized antibodies. Human antibodies in the form of scFvs expressed in the surface of these cells have been successfully diversified to produce a scFv library, from which …
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The functions of the RNA polymerase II CTD in transcription and RNA processing
… events, I created an Rpb1 conditional knock-out DT40 cell line (DT40-Rpb1) to further study the CTD with an initial focus on Thr 4, the function of which was unclear. Using DT40-Rpb1 system, we found that Thr 4 was phosphorylated in yeast, fly, chicken, and human cells, and cyclin-dependent …
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Characterising the function of CDK5RAP2 in the vertebrate centrosome
… motif 1 and 2. Using the chicken B-cell line, DT40, I have used gene-targeting methods to disrupt both of these domains in CDK5RAP2. This revealed a function for CDK5RAP2 in attaching centrosomes to mitotic spindle poles. Centrosome attachment to spindle poles is mediated by a binding partner …
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Characterization of the Tetraspanin protein Tspan18
… responds to calcium and MAPK signalling. In DT40 B cell lines, Tspan18 signalling was independent of Lyn, Syk, PLC2 and IP3 receptors, which are essential for B cell receptor signalling. However, Tspan18 signalling did require extracellular calcium and the calcium-activated phosphatase …
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An investigation into the regulation of replication factories and dormant origins in human cells
… show a role for RNF4 in factory regulation. In DT40 cells lacking RNF4, or in U2OS cells treated with RNF4 RNAi, factory regulation is abolished in response to stress. I also show that cells lacking dormant origins are hyper-sensitive to the effect of interstrand crosslinkers suggesting an …
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Towards the understanding of pericentriolar satellite biology
… determined the proteomic composition of PS from DT40 cells, I then performed validation studies both in chicken and human cell lines. In the second part of my study, I aimed to use the list of PS proteins to uncover new biological roles for pericentriolar satellites. I devised two distinct …
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Genetic interactions of repriming and translesion synthesis
… of G-quadruplex structures in the avian DT40 cell line. The growth inhibition of REV1-/- cells upon PhenDC3 treatment occurred in the apparent absence of DNA damage signalling pathway activation and was relieved upon knockout of p53, highlighting key differences in the response of …
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The Non-Wnt Functions of APC: Unravelling the Link between APC and Apoptosis
… APC. To this end, I created degron-tagged APC in DT40 cells that allowed for the rapid, conditional degradation of endogenous APC. The aim was to identify the immediate effects on cellular processes. Then, to identify the contribution of different APC domains by measuring the ability to rescue any …
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The role of the histone variant H3.3 and its chaperones in the response to DNA damage
… progression in the transformed avian cell line DT40, despite the availability during replication of canonical H3. In this thesis, I have examined the role of H3.3 and its chaperones in the response to UV induced DNA damage in a non-transformed, but immortalised, human cell system, the …
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The effect of replication impediments on differentiation
… impediments in the terminally differentiated DT40 chicken cell line. I wanted to find out whether these impediments could impact the fundamental biological process of embryonic development. In this work the BOBSC human induced pluripotent stem cell line was differentiated to definitive …
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The role of the BARD1 BRCT domains in the DNA damage response
… assays and colony survival assays, Bard1 DT40 cells reconstituted with BARD1?BRCT showed a significant decrease in their ability to carry out HR and were also extremely sensitive to PARP inhibitors suggesting that the BARD1 BRCT domains do indeed play an important role in HR. Work carried …
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Establishing the role of RNF4 in the vertebrate DNA damage response
… <br/> <br/>A homozygous knockout of RNF4 in the DT40 chicken lymphocyte cell line was generated to investigate the involvement of vertebrate RNF4 in protecting cells against DNA damage. Although the complete loss of RNF4 did not affect cell proliferation or cell cycle distribution, the RNF4 -/- …
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The effect of environmental stress and selective glucocorticoid receptor modulators on chicken and human leukaemia cells
… as well as chicken derived leukemia cells DT40. Compound B and C showed growth stimulatory effects and were not studied further. Leukaemia cells proliferation was mostly inhibited by high doses and long incubation time, whereas combination of compound treatment with either high or low …
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Microrna-mediated regulation and the fragile X family of proteins
… in a conditional Dicer knockdown cell line, DT40. We found that FXR1P, but not FMRP, increases upon Dicer knockdown and consequent absence of miRNAs suggesting that FXR1P is regulated by miRNAs, while FMRP is not. Expression of a luciferase reporter bearing the FXR1 3’UTR was significantly …
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Genetic selection of cyclic peptide G-quadruplex ligands
… we carried out such a genetic selection in DT40 cells, assessing expression loss of a surface protein BU-1 produced by an endogenous G4-containing locus. Peptides identified that disrupt BU-1 expression were then validated against a cell line with the BU-1 G4 edited out, allowing validation …
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Charakterisierung des Zytokins BAFF als wichtiger Regulator der B-Zellfunktion beim Haushuhn
… of chBAFF binding to the chicken B-cell line DT40 showed that an engineered soluble form of the human BAFF-receptor BCMA (huBCMA-Fc) inhibited chBAFF binding. The addition of 293T cell derived chBAFF to splenic lymphocyte cultures led to a significant increase in B-cell viability. This …