Copy Number Determination of Genetically-Modified Hematopoietic Stem Cells
Human gene transfer with gammaretroviral, murine leukemia virus (MLV) based vectors has been shown to effectively insert and express transgene sequences at a level of therapeutic benefit. However, there are numerous reports of disruption of the normal cellular processes caused by the viral insertion, even of replication deficient gammaretroviral vectors. Current gammaretroviral and lentiviral vectors do not control the site of insertion into the genome, hence, the possibility of disruption of the target cell genome. Risk related to viral insertions is linked to the number of insertions of the transgene into the cellular DNA, as has been demonstrated for replication competent and replication deficient retroviruses in experiments. At high number of insertions per cell, cell transformation due to vector induced activation of proto-oncogenes is more likely to occur, in particular since more than one transforming event is needed for oncogenesis. Thus, determination of the vector copy number in bulk transduced populations, individual colony forming units, and tissue from the recipient of the transduced cells is an increasingly important safety assay and has become a standard, though not straightforward assay, since the inception of quantitative PCR.
- 粘合连接(adheringjunction)
- Scalable Production of Embryonic Stem Cell-Derived Cells
- Electroporation of ES cells
- In Vitro Construction of 2D and 3D Simulations of the Murine Hematopoietic Niche
- Single-Cell Enzymatic Dissociation of hESC Lines OxF1OxF4 and Culture in Feeder-Free Conditions
- Single-Cell and Subcellular Measurement of Intracellular Ca2+ Concentration
- Generation of a Vascular Niche for Studying Stem Cell Homeostasis
- Gene Expression in Polytene Nuclei
- 脂锚定蛋白(lipid-anchored)
- Isolation of DNA Structure-Dependent Checkpoint Mutants in S. pombe