Generation of induced pluripotent stem cells by using a mammalian artificial chromosome expression system

Tóth, Ágota; Fodor, Katalin; Blazsó, Péter; Cserpán, Imre; Praznovszky, Tünde; Tubák, Vilmos; Udvardy, Andor; Hadlaczky, Gyula; Katona, Róbert L.

doi:10.1556/abiol.65.2014.3.9

Cited by 2 publications

(3 citation statements)

References 39 publications

(41 reference statements)

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“…Satellite-DNA-based artificial chromosomes (SATACs) have already passed the obstacles, including large-scale purification, transfer into various cells and embryos, germ-line transmission and generation of transgenic animals [90]. The reprogramming of MEFs was efficiently induced by HACs with engineered OSKM factors carrying an N-terminal flag-tag and a C-terminal polyarginine tail [91].…”

Section: Artificial Chromosome Systemmentioning

confidence: 99%

Current Reprogramming Systems in Regenerative Medicine: From Somatic Cells to Induced Pluripotent Stem Cells

2015

Regen. Med.

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Induced pluripotent stem cells (iPSCs) paved the way for research fields including cell therapy, drug screening, disease modeling and the mechanism of embryonic development. Although iPSC technology has been improved by various delivery systems, direct transduction and small molecule regulation, low reprogramming efficiency and genomic modification steps still inhibit its clinical use. Improvements in current vectors and the exploration of novel vectors are required to balance efficiency and genomic modification for reprogramming. Herein, we set out a comprehensive analysis of current reprogramming systems for the generation of iPSCs from somatic cells. By clarifying advantages and disadvantages of the current reprogramming systems, we are striding toward an effective route to generate clinical grade iPSCs.

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Section: Artificial Chromosome Systemmentioning

confidence: 99%

Current Reprogramming Systems in Regenerative Medicine: From Somatic Cells to Induced Pluripotent Stem Cells

2015

Regen. Med.

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“…Such research has included studies on the potential of HACs for gene function analysis, cell reprogramming and animal transgenesis. 1 − 7 , 12 − 22 …”

mentioning

confidence: 99%

“…Over the past 20 years, many groups have reported the successful generation of HACs of both types − (and references therein), and importantly all HACs constructed so far have constitutive kinetochores. , In the past few years, the amount of research based on the HAC-vectors has significantly increased due to the engineering of HACs with a single loxP gene integration site ,, (and references therein). Such research has included studies on the potential of HACs for gene function analysis, cell reprogramming and animal transgenesis. − ,− …”

mentioning

confidence: 99%

Method to Assemble Genomic DNA Fragments or Genes on Human Artificial Chromosome with Regulated Kinetochore Using a Multi-Integrase System

et al. 2017

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The production of cells capable of carrying multiple transgenes to Mb-size genomic loci has multiple applications in biomedicine and biotechnology. In order to achieve this goal, three key steps are required: (i) cloning of large genomic segments; (ii) insertion of multiple DNA blocks at a precise location and (iii) the capability to eliminate the assembled region from cells. In this study, we designed the iterative integration system (IIS) that utilizes recombinases Cre, ΦC31 and ΦBT1, and combined it with a human artificial chromosome (HAC) possessing a regulated kinetochore (alphoidtetO-HAC). We have demonstrated that the IIS-alphoidtetO-HAC system is a valuable genetic tool by reassembling a functional gene from multiple segments on the HAC. IIS-alphoidtetO-HAC has several notable advantages over other artificial chromosome-based systems. This includes the potential to assemble an unlimited number of genomic DNA segments; a DNA assembly process that leaves only a small insertion (<60 bp) scar between adjacent DNA, allowing genes reassembled from segments to be spliced correctly; a marker exchange system that also changes cell color, and counter-selection markers at each DNA insertion step, simplifying selection of correct clones; and presence of an error proofing mechanism to remove cells with misincorporated DNA segments, which improves the integrity of assembly. In addition, the IIS-alphoidtetO-HAC carrying a locus of interest is removable, offering the unique possibility to revert the cell line to its pretransformed state and compare the phenotypes of human cells with and without a functional copy of a gene(s). Thus, IIS-alphoidtetO-HAC allows investigation of complex biomedical pathways, gene(s) regulation, and has the potential to engineer synthetic chromosomes with a predetermined set of genes.

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Generation of induced pluripotent stem cells by using a mammalian artificial chromosome expression system

Cited by 2 publications

References 39 publications

Current Reprogramming Systems in Regenerative Medicine: From Somatic Cells to Induced Pluripotent Stem Cells

Current Reprogramming Systems in Regenerative Medicine: From Somatic Cells to Induced Pluripotent Stem Cells

Method to Assemble Genomic DNA Fragments or Genes on Human Artificial Chromosome with Regulated Kinetochore Using a Multi-Integrase System

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