Mammalian Artificial Chromosomes and Clinical Applications for Genetic Modification of Stem Cells: An Overview
Modifying multipotent, self-renewing human stem cells with mammalian artificial chromosomes (MACs), present a promising clinical strategy for numerous diseases, especially ex vivo cell therapies that can benefit from constitutive or overexpression of therapeutic gene(s). MACs are nonintegrating, autonomously replicating, with the capacity to carry large cDNA or genomic sequences, which in turn enable potentially prolonged, safe, and regulated therapeutic transgene expression, and render MACs as attractive genetic vectors for “gene replacement” or for controlling differentiation pathways in progenitor cells. The status quo is that the most versatile target cell would be one that was pluripotent and self-renewing to address multiple disease target cell types, thus making multilineage stem cells, such as adult derived early progenitor cells and embryonic stem cells, as attractive universal host cells. We will describe the progress of MAC technologies, the subsequent modifications of stem cells, and discuss the establishment of MAC platform stem cell lines to facilitate proof-of-principle studies and preclinical development.
- Hot Start PCR
- Down-Regulation of Gene Expression by RNA-Induced Gene Silencing
- Assembling Linear DNA Templates for In Vitro Transcription and Translation
- Motif-Dependent Polymerase Chain Reaction (PCR): DNA Fingerprinting Enterotoxigenic Escherichia coli
- Cryopreservation of Transgenic Sheep Lines
- Mutagen: A Random Mutagenesis Method Providing a Complementary Diversity Generated by Human Error-Prone DNA Polymerases
- Conditional Gene-Trap Mutagenesis in Zebrafish
- DNA Extraction of Ancient Animal Hard Tissue Samples via Adsorption to Silica Particles
- Fluorochrome-Labeled Inhibitors of Caspases: Convenient In Vitro and In Vivo Markers of Apoptotic Cells for Cytometric Analysis
- Positional Cloning and Multicolor In Situ Hybridization Principles and Protocols