TY - JOUR
T1 - Generation of induced pluripotent stem cells using elastin like polypeptides as a non-viral gene delivery system
AU - Lee, Chang Hyun
AU - Ingrole, Rohan S.J.
AU - Gill, Harvinder Singh
N1 - Publisher Copyright:
© 2019 Elsevier B.V.
PY - 2020/4/1
Y1 - 2020/4/1
N2 - Induced pluripotent stem cells (iPSCs) have been generated from various somatic cells using different approaches; however, a major restriction of reprogramming methods is the use of viral vectors, which have the risk of causing genome-integration of viral DNA. Here, without a viral vector, we generated iPSCs from mouse fibroblasts using an elastin-like polypeptide (ELP)-based transfection method. Our findings support the possible use of ELPs for delivery of the reprogramming genes in to somatic cells for generation of iPSCs. Results of gel retardation assay demonstrated efficient complexation of ELPs with a plasmid containing the four Yamanaka stem cell factors, Oct-4, Klf4, c-myc, and Sox2. After transfection, the iPSCs showed embryonic stem cell-like characteristics, including expression of endogenous pluripotency genes, differentiation into three germ layer lineages, and formation of teratomas in vivo. Our results demonstrate that ELP-based gene delivery may provide a safe method for use in generation of virus-free and exogenous DNA-free iPSCs, which will be crucial for future applications in stem cell-based therapies.
AB - Induced pluripotent stem cells (iPSCs) have been generated from various somatic cells using different approaches; however, a major restriction of reprogramming methods is the use of viral vectors, which have the risk of causing genome-integration of viral DNA. Here, without a viral vector, we generated iPSCs from mouse fibroblasts using an elastin-like polypeptide (ELP)-based transfection method. Our findings support the possible use of ELPs for delivery of the reprogramming genes in to somatic cells for generation of iPSCs. Results of gel retardation assay demonstrated efficient complexation of ELPs with a plasmid containing the four Yamanaka stem cell factors, Oct-4, Klf4, c-myc, and Sox2. After transfection, the iPSCs showed embryonic stem cell-like characteristics, including expression of endogenous pluripotency genes, differentiation into three germ layer lineages, and formation of teratomas in vivo. Our results demonstrate that ELP-based gene delivery may provide a safe method for use in generation of virus-free and exogenous DNA-free iPSCs, which will be crucial for future applications in stem cell-based therapies.
KW - ELPs
KW - Elastin like proteins
KW - Gene delivery
KW - Non-viral gene delivery
KW - iPSC generation
UR - http://www.scopus.com/inward/record.url?scp=85062018166&partnerID=8YFLogxK
U2 - 10.1016/j.bbadis.2019.01.031
DO - 10.1016/j.bbadis.2019.01.031
M3 - Article
C2 - 30753882
AN - SCOPUS:85062018166
SN - 0925-4439
VL - 1866
JO - Biochimica et Biophysica Acta - Molecular Basis of Disease
JF - Biochimica et Biophysica Acta - Molecular Basis of Disease
IS - 4
M1 - 165405
ER -