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Pluripotent stem cell-based gene therapy approach: human de novo synthesized chromosomes

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Abstract

A novel approach in gene therapy was introduced 20 years ago since artificial non-integrative chromosome-based vectors containing gene loci size inserts were engineered. To date, different human artificial chromosomes (HAC) were generated with the use of de novo construction or “top-down” engineering approaches. The HAC-based therapeutic approach includes ex vivo gene transferring and correction of pluripotent stem cells (PSCs) or highly proliferative modified stem cells. The current progress in the technology of induced PSCs, integrating with the HAC technology, resulted in a novel platform of stem cell-based tissue replacement therapy for the treatment of genetic disease. Nowadays, the sophisticated and laborious HAC technology has significantly improved and is now closer to clinical studies. In here, we reviewed the achievements in the technology of de novo synthesized HACs for a chromosome transfer for developing gene therapy tissue replacement models of monogenic human diseases.

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Abbreviations

BACs:

Bacterial artificial chromosomes

CENP-B:

Centromere protein B

cGMP:

Current good manufacturing practice

GFP:

Green fluorescent protein

CHO:

Chinese hamster ovary

ESCs:

Embryonic stem cells

IIS:

Iterative integration system

iMCT:

Isolated metaphase chromosome transfection

iPSCs:

Induced pluripotent stem cells

FVIII:

Human clotting factor VIII

HAC:

Human artificial chromosome

hESCs:

Human embryonic stem cells

hiPSCs:

Human induced pluripotent stem cells

HPRT:

Hypoxanthine–guanine phosphoribosyl transferase

HSV-1:

Herpes simplex virus 1

HSPCs:

Hematopoietic stem and progenitor cells

HVJ-E:

Hemagglutinating virus of Japan E

lacO:

Lac operon

loxP:

Locus of X-over P1

MLV:

Murine leukemia retrovirus

MMCT:

Microcell mediated chromosome transfer

MSCs:

Mesenchymal stem cells

MV:

Measles virus

PEG:

Polyethylene glycol

PSCs:

Pluripotent stem cells

rAAVs:

Adeno-associated recombinant viruses

RMCE:

Recombinase-mediated cassette exchange method

TAR:

Transformation associated recombination

tetO:

Tetracycline operator

tetR:

Tet-repressor protein

SIM:

Sequential integration of multiple vectors

YACs:

Yeast artificial chromosomes

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Acknowledgements

We thank the reviewer for valuable comments. This work was supported by the Saint-Petersburg State University intramural grant 60257027, Russian Science Foundation grant 20-14-00242.

Funding

The Saint-Petersburg State University intramural grant 60257027, Russian Science Foundation grant 20–14-00242.

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  1. Institute of Cytology, Russian Academy of Sciences, 4 Tikhoretsky Ave, St-Petersburg, 194064, Russia

    Sergey A. Sinenko, Sergey V. Ponomartsev & Alexey N. Tomilin

  2. Institute of Translational Biomedicine, St-Petersburg State University, 7–9, Universitetskaya Emb, St-Petersburg, 199034, Russia

    Alexey N. Tomilin

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  1. Sergey A. Sinenko
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S.A.S.: writing a draft of the manuscript; S.V.P.: figure artwork; S.A.S., S.V.P. and A.N.T.: manuscript writing; A.N.T.: manuscript review and editing; S.A.S. and A.N.T.: review and final approval of the manuscript.

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Sinenko, S.A., Ponomartsev, S.V. & Tomilin, A.N. Pluripotent stem cell-based gene therapy approach: human de novo synthesized chromosomes. Cell. Mol. Life Sci. 78, 1207–1220 (2021). https://doi.org/10.1007/s00018-020-03653-1

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