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Using inositol as a biocompatible ligand for efficient transgene expression.

Zhang L, Bellis SL, Fan Y, Wu Y - Int J Nanomedicine (2015)

Bottom Line: The three resulting PG6-PEI-INO polymers have an increased number of INO ligands per molecule.Studies further revealed that extracellular adenosine triphosphate (eATP) can inhibit the transgene efficiency of PG6-PEI-INO polymers, as compared with PEI and PG6-PEI that were not conjugated with inositol.Our work unveiled the possibility of using inositol as an effective ligand for transgene expression.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, People's Republic of China.

ABSTRACT
Transgene transfection techniques using cationic polymers such as polyethylenimines (PEIs) and PEI derivatives as gene vectors have shown efficacy, although they also have shortcomings. PEIs have decent DNA-binding capability and good cell internalization performance, but they cannot deliver gene payloads very efficiently to cell nuclei. In this study, three hyperbranched polyglycerol-polyethylenimine (PG6-PEI) polymers conjugated with myo-inositol (INO) molecules were developed. The three resulting PG6-PEI-INO polymers have an increased number of INO ligands per molecule. PG6-PEI-INO 1 had only 14 carboxymethyl INO (CMINO) units per molecule. PG6-PEI-INO 2 had approximately 130 CMINO units per molecule. PG6-PEI-INO 3 had as high as 415 CMINO units approximately. Mixing PG6-PEI-INO polymers with DNA produced compact nanocomposites. We then performed localization studies using fluorescent microscopy. As the number of conjugated inositol ligands increased in PG6-PEI-INO polymers, there was a corresponding increase in accumulation of the polymers within 293T cell nuclei. Transfection performed with spherical 293T cells yielded 82% of EGFP-positive cells when using PG6-PEI-INO 3 as the vehicle. Studies further revealed that extracellular adenosine triphosphate (eATP) can inhibit the transgene efficiency of PG6-PEI-INO polymers, as compared with PEI and PG6-PEI that were not conjugated with inositol. Our work unveiled the possibility of using inositol as an effective ligand for transgene expression.

No MeSH data available.


Synthesis route of PG6-PEI-INO polymers.Abbreviations: CMPG6, carboxymethyl polyglycerol; CMINO, carboxymethyl inositol; DCC, N,N’-dicyclohexylcarbodiimide; DMF, N-dimethyl-formamide; EDC, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydro chloride; INO, myo-inositol; NHS, N-hydroxysuccinimide; PEI, polyethylenimine; PG6, polyglycerol.
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f2-ijn-10-2871: Synthesis route of PG6-PEI-INO polymers.Abbreviations: CMPG6, carboxymethyl polyglycerol; CMINO, carboxymethyl inositol; DCC, N,N’-dicyclohexylcarbodiimide; DMF, N-dimethyl-formamide; EDC, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydro chloride; INO, myo-inositol; NHS, N-hydroxysuccinimide; PEI, polyethylenimine; PG6, polyglycerol.

Mentions: GPC (SEC) analysis showed that PG6-PEI-INO 1, 2, and 3 (Figure 2) had weight average molecular weights (Mw) of 455, 465, and 542 kDa, respectively (Table 1).


Using inositol as a biocompatible ligand for efficient transgene expression.

Zhang L, Bellis SL, Fan Y, Wu Y - Int J Nanomedicine (2015)

Synthesis route of PG6-PEI-INO polymers.Abbreviations: CMPG6, carboxymethyl polyglycerol; CMINO, carboxymethyl inositol; DCC, N,N’-dicyclohexylcarbodiimide; DMF, N-dimethyl-formamide; EDC, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydro chloride; INO, myo-inositol; NHS, N-hydroxysuccinimide; PEI, polyethylenimine; PG6, polyglycerol.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4403686&req=5

f2-ijn-10-2871: Synthesis route of PG6-PEI-INO polymers.Abbreviations: CMPG6, carboxymethyl polyglycerol; CMINO, carboxymethyl inositol; DCC, N,N’-dicyclohexylcarbodiimide; DMF, N-dimethyl-formamide; EDC, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydro chloride; INO, myo-inositol; NHS, N-hydroxysuccinimide; PEI, polyethylenimine; PG6, polyglycerol.
Mentions: GPC (SEC) analysis showed that PG6-PEI-INO 1, 2, and 3 (Figure 2) had weight average molecular weights (Mw) of 455, 465, and 542 kDa, respectively (Table 1).

Bottom Line: The three resulting PG6-PEI-INO polymers have an increased number of INO ligands per molecule.Studies further revealed that extracellular adenosine triphosphate (eATP) can inhibit the transgene efficiency of PG6-PEI-INO polymers, as compared with PEI and PG6-PEI that were not conjugated with inositol.Our work unveiled the possibility of using inositol as an effective ligand for transgene expression.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, People's Republic of China.

ABSTRACT
Transgene transfection techniques using cationic polymers such as polyethylenimines (PEIs) and PEI derivatives as gene vectors have shown efficacy, although they also have shortcomings. PEIs have decent DNA-binding capability and good cell internalization performance, but they cannot deliver gene payloads very efficiently to cell nuclei. In this study, three hyperbranched polyglycerol-polyethylenimine (PG6-PEI) polymers conjugated with myo-inositol (INO) molecules were developed. The three resulting PG6-PEI-INO polymers have an increased number of INO ligands per molecule. PG6-PEI-INO 1 had only 14 carboxymethyl INO (CMINO) units per molecule. PG6-PEI-INO 2 had approximately 130 CMINO units per molecule. PG6-PEI-INO 3 had as high as 415 CMINO units approximately. Mixing PG6-PEI-INO polymers with DNA produced compact nanocomposites. We then performed localization studies using fluorescent microscopy. As the number of conjugated inositol ligands increased in PG6-PEI-INO polymers, there was a corresponding increase in accumulation of the polymers within 293T cell nuclei. Transfection performed with spherical 293T cells yielded 82% of EGFP-positive cells when using PG6-PEI-INO 3 as the vehicle. Studies further revealed that extracellular adenosine triphosphate (eATP) can inhibit the transgene efficiency of PG6-PEI-INO polymers, as compared with PEI and PG6-PEI that were not conjugated with inositol. Our work unveiled the possibility of using inositol as an effective ligand for transgene expression.

No MeSH data available.