International Journal of Pharmaceutical Sciences and Nanotechnology
Aggarwal et al: Polyplex: A Promising Gene Delivery System
Review Article
Polyplex: A Promising Gene Delivery System
Rohan Aggarwal*, Monika Targhotra, Bhumika Kumar, P.K Sahoo and Meenakshi
K. Chauhan
Department of Pharmaceutics, Delhi Institute of Pharmaceutical Science and Research, New
Delhi-110017, India.
Received September 19, 2019; accepted October 7, 2019
ABSTRACT
In the past few years gene delivery system has gained a
huge attention owing to its proved efficacy in several
diseases especially in those caused by genetic and/or
oncological malfunctioning. The effective gene delivery
mainly depends on the carrier molecules that can ensure
the safe and specific delivery of the nucleic acid
molecules. Viral vectors have been used for a longer
period as the gene transfer vehicle. However, these viral
vectors have potential immunological disadvantages that
made them less preferred. Recently, non-viral vectors
such as polyplexes have emerged as a promising
alternative for viral vectors. Polyplexes are formed by
KEYWORDS: Polyplex; Gene delivery; Structure; Stability; Polyethyleneimine; Cytotoxicity.
Introduction
Gene therapy mainly includes delivery of nucleic acid
and therapeutics that are frequently used to attune the
cellular expression of some peculiar targeted genes along
with their proteins to treat hereditary as well as
acquired diseases (Chakraborty et al., 2017). Vectors play
important role in gene therapy since these are the only
means to deliver specific gene inside cell. Vectors may be
viral and non-viral as classified in figure 1. Non-viral
vectors are further classified into two groups like
chemical and physical vector. Chemical vectors include
poly-cationic carriers such as liposomes (lipoplexes) and
polymers (polyplexes).
Fig. 1. Vector classification.
Events in gene transfection process of non-viral
vector is similar to the viral vector. The events are
represented in Figure 2.
After administration non-viral vector encounters
various systemic as well as cellular barriers are
represented in Figure 3.
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Volume 12 • Issue 6 • November – December 2019
MS ID: IJPSN-9-20-19-AGGARWAL
conjugating a polymer with DNA and in maximum cases
the cationic polymers are preferred over others. The
structure and stability of the polyplexes depends on
various factors. The ability of the polymer to condense
the DNA mainly dictates the efficiency of the polyplex
mediated transfection. In this review we are going to
provide a framework for the synthesis and design of the
polyplexes along with the structure and stability of the
complexes pertaining to mechanism of action,
characterization and therapeutic application, including
polyethyleneimine mediated cytotoxicity as well as newer
strategies for the generation of better polyplexes.
Formation of nucleic acid-polymer complex by
combining the nucleic acids with non-viral polymeric
carriers via electrostatic interactions is commonly called
as polyplexes (Lächelt and Wagner, 2015). A huge
number of cationic polymers used as nucleic acid
carriers, like polyethylenimine (PEI), poly-L-lysine
(PLL), polyamidoamine (PAMAM), and poly (2-
dimethylaminoethyl methacrylate) (PDMAEMA)
(Agarwal et al., 2012).
This review addresses the formation of polyplexes
along with the structure and stability of the complexes
pertaining to mechanism of action, characterization and
therapeutic application, including polyethyleneimine
mediated cytotoxicity as well as newer strategies for the
generation of better polyplexes.
Composition
Cationic polyplexes are formed in various lengths,
with various geometrical status and exchange or
inclusion of different functional groups in a relatively
flexible procedure (Elouahabi and Ruysschaert, 2005).
Polyplex formation is mainly controlled by the kinetics.
The formation should conduct at the particular ionic
strengths where the polycation formation is fast and
permanent. The resulting polyplex size and the
transfection capacity can be influenced by the order of
incorporation of reacting substances throughout the
complexation process (Tros et al., 2010).
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4682
Fig. 2. Events in non-viral transfection.
Fig. 3. Systemic and cellular barriers of non-viral vectors.
According to a study, high molecular-weight
polyethyleneimine condensed plasmid DNA in the same
way as compare with low-molecular-weight polyethyle-
neimine in the same Negative/ Positive charge ratio
(Kunath et al., 2003). Another study estimated that the
polyplex size synthesized between DNA and branched
25-kDa polyethyleneimine or linear 22-kDa polyethy-
leneimine may range from 20 to 40 nm when the
Negative/ Positive charge ratio was 1.6. The study
further added that complete DNA condensation observed
at this ratio (Elouahabi and Ruysschaert, 2005). Results
from photon correlation spectroscopy reported that
polyplexes formed between molecular weight of 25 kDa
polyethyleneimine and DNA at a Negative/ Positive
charge ratio of 6.7 showed a size of almost 150 nm
(Kunath et al., 2003).
Several studies reported that to produce stable
polyplex complexes, using branched (Petersen et al.,
2002) or linear polyethyleneimine (Jeong et al., 2001). A
negative/positive charge ratio of about 2–3 is necessary.
Another study suggested that DNA complexation with
different sized polyethyleneimine had not been shown to
result for an alteration of DNA conformation, which
persisted specifically in B form. The study further added
that low ionic strength enhanced the changes of
protonation in polyethylenimine, therefore, possible
shifts in pKa of ionizable groups of polyethylenimine as a
result of DNA complexation suggested. The size and zeta
potential of polyethylenimine and DNA polyplex was not
influenced by the molecular weight or the linearity or
branching of polyethylenimine. Whereas, they depend
mainly on the negative/positive charge ratio used for the
complexation procedure (Choosakoonkriang et al., 2003).
It is reported that, some sodium salts like sodium
phosphate showed better compatibility than any other
buffer system like NaCl or phosphate-buffered saline in
in vivo administration process (Hartikka et al., 2000). It
Int J Pharm Sci Nanotech Vol 12; Issue 6 • November− December 2019
is reported that the DNA is usually completely bound
and compacted at a charge ratio of 1 or above 1. The
study further added that polyplexes may aggregate with
lower solubility around electroneutrality, however, it was
found that a large proportion of the polymer was not
bound to DNA at high charge ratios (Wagner, 2004).
Structure and Stability
The tough electrostatic reactions between
polyethyleneimine and DNA transform the DNA in a
condensed form and therefore a nanostructured
positively charged complex called polyplex generated.
Fourier transform infrared (FTIR) data explains the
electrostatic bonding between DNA and the polymer
explicitly. A depletion in the frequency of the phosphate
stretching vibration of plasmid DNA is reported after
having complex reaction with polyethyleneimine
(Choosakoonkriang et al., 2003). Complexation behavior
is influenced by the degree of polymer division. The study
showed that smaller polyplexes originated from
hyperbranched polyethyleneimine complexes with
secondary and tertiary amine have higher transfection
efficiency (Banerjee et al., 2004). The capacity of DNA
condensation for linear polyethyleneimine is less owing
to its minimum content of primary amines compared to
the branched forms (Tros et al., 2010).
Researchers found that polyethyleneimine and DNA
alone can freely soluble in water, but polyplex of
polyethyleneimine and DNA is insoluble in water. Final
structure and transfection efficiency of polyplexes are
largely dependent upon storage condition (Banerjee
et al., 2004). Transfection capacity of the polyplexes is
affected by the complexation, size, and structural
properties. Therefore, higher branched polymer can
make better DNA condensation and provide better
protection against degradation in extracellular
environment. Finally, cell uptake improved by the
Aggarwal et al: Polyplex: A Promising Gene Delivery System
electrostatic interactions of the polycation with the
negatively charged cellular surface.
Mechanism of Action
Cell Binding and Uptake Mechanism
It is reported that cationic polyplexes can enter the
cell through interaction with the transmembrane protein
called syndecan (Kunath et al., 2003). Several studies
showed that the internalization of polyplexes via
transfection process was depended upon the type of cell
and the polymer. It is reported that for the linear
polyplexes transfection occurred via clathrin-coated pit
pathway. In contrast, in the transfection of branched
polyplexes, both clathrin-mediated and the lipid raft
pathways are involved. Therefore, HeLa cells were
dependent upon both the lipid-raft and clathrin-mediated
pathways (Gersdorff et al., 2006). Whereas, another
study revealed that in COS-7 cells, both the clathrin- and
caveolae-dependent pathways involved in internalization
process (vander et al., 2007). According to a different
study in HeLa and A549 cells, polyplexes were
incorporated in cells via caveolae pathway and had
complete transection (Rejman et al., 2006).
Endosomal Escape of Polyplexes
Several studies assumed that after cellular
internalization, endosomal escape and high transfection
capacity of polyplexes can be explained by the hypotheses
called “proton-sponge effect” (Taranejoo et al., 2015;
Venkiteswaran et al., 2016). According to the
hypothesis, 1–6 nitrogen atoms are protonated at
physiological pH while in the acidic endosomal pH, the
number of protonated nitrogen elevated, and produces a
charge gradient thereby a Cl− influx induced. Therefore,
water influx induced by the increased Cl− concentration,
and, finally endosome swelling leads to rupture of
polyplexes and releasing the components into the
cytoplasm. Another study reported that usually for
polymers with an acidic buffer (pH -5) capacity may not
be suitable for explaining the proton sponge hypothesis
(Tros et al., 2010).
Mechanism of polyplexes dissociation
After the endosomal escape, dissociation is the
important stage for the polyplexes. Low molecular-
weight Poly-L-Lysine-based polyplexes have high
dissociation rate and more transfection efficacy than high
molecular weight polyplexes. However, polyethylenei-
mine containing polyplexes have relatively slower
dissociation rate (Ogris et al., 2001). A study reported
that after 18 hours of transfection, pancreatic carcinoma
cell line with double-fluorescence-labeled polyethylenei-
mine-DNA polyplexes have shown few condensed
polyplexes and many dissociated polyethyleneimine
polyplex (Bieber et al., 2002).
Nuclear import
Post dissociation, to provide DNA preservation or
cytoplasmic movability nuclear import of DNA may
proceed through indirect procedures (Lam and dean,
4683  
2010). However, it is also reported that when DNA and
polyethyleneimine were directly labeled, they remained
in complex form inside the cell and localized to the
nucleus coordinately. Whereas, other study suggested
that, in the time of the cell division when the nuclear
membrane is dismantled for some time a passive way is
used for plasmid DNA entry into the cell nucleus
(Männistö et al., 2007).
Characterization
A number of characterization techniques can
demonstrate the structure of DNA-polyplex complexes to
understand transfection efficacy. There are different
light scattering tools that are used for characterization of
the colloidal features of the polyplexes. Fluorescence
techniques are used frequently to study the DNA
condensation. For evaluation of topographical description
of the polyplexes electron and scanning probe microscopy
are used. Nuclear magnetic resonance (NMR) and
electron spin resonance (ESR) spectroscopy are used to
determine the structural properties of particular polymer
segments (Tros et al., 2010). Laser scanning confocal
microscopy (LSCM) can analyze the activities of
complexes inside the cells and the changes happening
following DNA release. Quantitative structure-activity
relationship (QSAR) modeling can examine the molecular
basis of gene delivery (Horobin and Weissig, 2005). Gel
permeation chromatography (GPC) is used to determine
the cloud point (CP) of thermosensitive polymers.
Dynamic light scattering (DLS) can analyze the
hydrodynamic size and polydispersity index (PDI) of the
polyplexes. Laser Doppler Electrophoresis (LDE) is used
to determine the ζ-potential of the polyplexes. Static light
scattering (SLS) is used to measure the gyration and
hydrodynamic radius of the polyplexes (Fliervoet et al.,
2019).
Researchers characterized siRNA polyplex with a
comparison of different particle sizing methods namely
DLS, atomic force microscopy (AFM), nanoparticle
trafficking analysis (NTA) and fluorescence correlation
spectroscopy (FCS). They reported that all the methods
were able to analyze larger than 120 nm polydisperse but
NTA was unable to measure smaller than 40 nm primary
particles (Troiber et al., 2013). It is reported that Taylor
dispersion analysis (TDA) was successfully used for the
size characterization of polyplexes and the quantification
of unbound polycation within the polyplex (Leclercq
et al., 2015).
Application
Gene therapy is a budding treatment option for
acquired diseases like AIDS and cancer along with
hereditary disorders, like cystic fibrosis and hemophilia
(Martinez-Fong et al., 2012). Neurotensin (NTS) receptor
type 1 (NTSR1) exhibit distinctive structural and
characteristics to treat inconsistent types of cancer cell.
Therefore, neurotensin (NT)-polyplex is a non-viral
targeted gene delivery system consists of synthetic
nanoparticles that can transfer plasmid DNA (pDNA)
encoding any specific targeted gene into cells that
4684 Int J Pharm Sci Nanotech Vol 12; Issue 6 • November− December 2019
express and internalize the NTSR1, like dopaminergic
neuron system (Katragadda et al., 2010) and cancer cells
(Rubio-Zapata et al., 2009).
The sympathetic nervous system may produce the
most common extracranial solid tumor called
Neuroblastoma. Therefore, NT-polyplex is a novel
therapeutic alternative of advanced staged
Neuroblastoma because of the selective, potent,
biodegradable and nontoxic properties (Arango-
Rodriguez et al., 2006; Gonzalez-Barrios et al., 2006).
According to a study, the NT polyplex could transfect
therapeutic genes into Neuroblastoma tumor cells across
the bloodstream administration or through intratumoral
injection. The study also reported that the thymidine
kinase (HSVTK) suicide gene transfection along with
Ganciclovir (GCV) treatment reduced the size and weight
of Neuroblastoma tumors by 30–50% and induced
apoptosis compared to controls (Rubio-Zapata et al.,
2009).
An aggressive form of breast cancer can be treated
with the targeted gene-mediated synthetic polyplexes
(Jemal et al., 2011). Studies have shown that targeted
therapy with therapeutic nanoparticles combined with a
specific ligand can be delivered to an over-expressed
receptor in breast cancer cells. The major target for this
therapy is the Human epidermal growth factor-2 (HER2)
receptor in the HER2-induced breast cancer. Cancer can
be treated with polyplexes containing a recombinant
humanized monoclonal antibody or a single-chain
antibody fragment as ligands (Huang et al., 2010).
The human breast adenocarcinoma cell line such as
MDA-MB-231 express neurotensin receptor type 1
(NTSR1) and thus making it a target of therapeutic
genes carried by neurotensin (NTS)-polyplex nanocarrier.
Recent study reported that NTS-polyplex nanoparticles
along with the Herpes simplex virus thymidine kinase
(HSVtk) suicide gene and Ganciclovir (GCV), transfected
both genes in cultured MDA-MB-231 cells. HSVtk gene
expression reduced cell viability by 49% and increased
apoptosis in cultured MDA-MB-231 cells after GCV
treatment (Castillo-Rodríguez et al., 2014).
A cumulative depletion of dopaminergic neurons in
the substantia nigra with contemporaneous gliosis in
midbrain may produce Parkinson’s disease. The NTS
carrier makes endocytosis of NTS with its high-affinity
receptor Neurotensin receptor type 1 to deliver the
plasmid DNA into dopamine neurons (Arango-Rodriguez
et al., 2006). According to a study, the NTS-polyplex used
a fusogenic peptide (FP) and a karyophilic peptide (KP)
to increase transfection efficiency (Esbjörner et al., 2007).
The fusogenic peptide is bound with the poly-lysine
moiety; to protect NTS-polyplex from endosomal uptake
before the enzymatic degradation. The karyophilic
peptide provides a nuclear localization signal which is
electrostatically linked to the plasmid DNA to assist
nuclear import.
Toxicity
It is considered that polyethyleneimine is a secure
carrier for gene-mediated therapy, but its cytotoxicity is
challenging for researchers (Ramamoorth, 2015).
Therefore, polyethyleneimine complexes had revealed
cell toxicity effects (Moghimi et al., 2005). Cell necrotic
and apoptotic pathways are also affected by the
cytotoxicity. The toxicity depends upon the molecular
weight, degree of branching, size and zeta potential of
derivatives prepared from polyethyleneimine. Mostly, the
higher molecular weight polymer and high ionic strength
buffer can produce more compact nanoparticles,
therefore, cytotoxicity can be affected. Several studies
have reported that the cytotoxicity is inversely related to
polyplex size, therefore, bigger polyplexes had lower
cytotoxicity and smaller polyplexes showed higher
cytotoxicity (Kunath et al., 2003).
A recent study investigated the polyplexes by
evaluating the changes in sizes of polyplexes through
molecular weight, cell medium and branching degree of
the polymer to evaluate their effects on the cytotoxicity.
The study showed that particles size was inversely
linked with molecular weight in case of branched
polyethyleneimine. According to this study the
cytotoxicity reports revealed that linear 250 kDa
polyethyleneimine was non-toxic whereas lower
molecular weight branched polyethyleneimine explained
toxicity effects in a concentration-dependent fashion. The
study also reported that the cytotoxicity effects of
branched polyethyleneimine were in proportional with
carrier/plasmid ratio and more significant for the
polyplexes prepared from HBG buffer. Results from Flow
cytometry analysis suggested that apoptosis is the major
procedure of polyplex induced cell toxicity (Kazemi et al.,
2018).
Future Prospects
Although polyethylenimine polyplexes comprise many
significant abilities required for effective nucleic acid
delivery, many cationic polymers with excellent
transfection properties are usually found to be cytotoxic
(Fischer et al., 2003). Numerous strategies have been
developed to reduce polyethyleneimine induced
cytotoxicity and increase the efficiency like size control
and topology, use of biodegradable cross-linked to low
molecular weight polyethyleneimine, use of modified
statistical surface and co-polymers and conjugation of
oligoamine segment (Hall et al., 2017).
To achieve better colloidal stability, reduced
cytotoxicity and relatively low immunological response
newer ternary complexes containing cationic lipids, a
cationic polymer, and nucleic acids are developed namely
Lipopolyplexes (LPPs) (Urbiola et al., 2013). Lipopoly-
plexes can combine the complexation properties of
cationic liposomes with cationic polymers, therefore,
DNA (Weide et al., 2008), siRNA (Li S-D et al., 2006), or
mRNA (Mockey et al., 2007) can be delivered.
Decationized polyplexes have also shown an excellent
safety profile, better stability, and improvement in vivo
circulation half-life and target tissue accumulation.
Therefore, the decationized polyplexes could be a great
choice for targeted gene-mediated therapies requiring
systemic administration (Novo et al., 2015).
Aggarwal et al: Polyplex: A Promising Gene Delivery System
Conclusions
Polyplexes are suggested as an excellent medium for
gene delivery without the serious immunological issues
associated with viral systems. Although newly developed
polyplexes appears to overcome many drawbacks, they
are also having certain limitations. Therefore, newer
approaches should meet quality demands like
effectiveness, toxicity, and feasibility to meet patient’s
criteria.
Acknowledgements
No institutional, private or corporate financial
support for the work was provided.
Conflict of interest
Conflict of interest declared none.
References
Agarwal S, Zhang Y, Maji S and Greiner A (2012). PDMAEMA based
gene delivery materials. Mater Today 15(9): 388-393.
Arango-Rodriguez ML, Navarro-Quiroga I, Gonzalez-Barrios JA,
Martinez-Arguelles DB, Bannon MJ and Kouri J (2006).
Biophysical characteristics of neurotensin polyplex for in vitro
and in vivo gene transfection. Biochim Biophys Acta 1760(7):
1009-1020.
Banerjee P, Reichardt W, Weissleder R and Bogdanov A (2004).
Novel Hyperbranched Dendron for Gene Transfer in Vitro and in
Vivo. Bioconjug Chem 15(5): 960-968.
Bieber T, Meissner W, Kostin S, Niemann A and Elsasser H-P
(2002). Intracellular route and transcriptional competence of
polyethylenimine-DNA complexes. J Control Release Off J
Control Release Soc 82(2-3): 441-454.
Castillo-Rodríguez RA, Arango-Rodríguez ML, Escobedo L,
Hernandez-Baltazar D, Gompel A and Forgez P (2014). Suicide
HSVtk Gene Delivery by Neurotensin-Polyplex Nanoparticles
via the Bloodstream and GCV Treatment Specifically Inhibit the
Growth of Human MDA-MB-231 Triple Negative Breast Cancer
Tumors Xenografted in Athymic Mice. PLoS ONE 9(5): e97151.
Chakraborty C, Sharma AR, Sharma G, Doss CGP and Lee S-S
(2017). Therapeutic miRNA and siRNA: Moving from Bench to
Clinic as Next Generation Medicine. Mol Ther - Nucleic Acids 8:
132-143.
Choosakoonkriang S, Lobo BA, Koe GS, Koe JG and Middaugh
CRussell (2003). Biophysical Characterization of PEI/DNA
Complexes. J Pharm Sci 92(8):1710-1722.
Elouahabi A and Ruysschaert J-M (2005). Formation and
Intracellular Trafficking of Lipoplexes and Polyplexes. Mol Ther
11(3): 336-347.
Esbjörner EK, Oglcka K, Lincoln P, Gräslund A and Nordén B
(2007). Membrane Binding of pH-Sensitive Influenza Fusion
Peptides. Positioning, Configuration, and Induced Leakage in a
Lipid Vesicle Model †. Biochemistry 46(47): 13490-13504.
Fischer D, Li Y, Ahlemeyer B, Krieglstein J and Kissel T (2003). In
vitro cytotoxicity testing of polycations: influence of polymer
structure on cell viability and hemolysis. Biomaterials 24(7):
1121-1131.
Fliervoet LAL, van Nostrum CF, Hennink WE and Vermonden T
(2019). Balancing hydrophobic and electrostatic interactions in
thermosensitive polyplexes for nucleic acid delivery. Multifunct
Mater 2(2): 024002.
Gonzalez-Barrios JA, Lindahl M, Bannon MJ, Anaya-Martínez V,
Flores G and Navarro-Quiroga I (2006). Neurotensin polyplex as
an efficient carrier for delivering the human GDNF gene into
nigral dopamine neurons of hemi parkinsonian rats. Mol Ther
14(6): 857-865.
4685  
Hall A, Lächelt U, Bartek J, Wagner E and Moghimi SM (2017).
Polyplex Evolution: Understanding Biology, Optimizing
Performance. Mol Ther 25(7):1476-1490.
Hartikka J, Bozoukova V, Jones D, Mahajan R, Wloch M and
Sawdey M (2000). Sodium phosphate enhances plasmid DNA
expression in vivo. Gene Ther 7(14): 1171-1182.
Horobin RW and Weissig V (2005). A QSAR-modeling perspective on
cationic transfection lipids. 1. Predicting efficiency and
understanding mechanisms. J Gene Med 7(8): 1023-1034.
Huang H, Yu H, Tang G, Wang Q and Li J (2010). Low molecular
weight polyethylenimine cross-linked by 2-hydroxypropyl-γ-
cyclodextrin coupled to peptide targeting HER2 as a gene
delivery vector. Biomaterials 31(7): 1830-1838.
Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D
(2011). Global cancer statistics. CA Cancer J Clin 61(2): 69-90.
Jeong JH, Song SH, Lim DW, Lee H and Park TG (2001). DNA
transfection using linear poly(ethylenimine) prepared by
controlled acid hydrolysis of poly(2-ethyl-2-oxazoline). J Control
Release Off J Control Release Soc 73(2-3): 391-399.
Katragadda CS, Choudhury PK and Murthy PN (2010).
Nanoparticles as non-viral gene delivery vectors. Indian J
Pharm Educ Res 44(2): 109-111.
Kazemi Oskuee R, Dabbaghi M, Gholami L, Taheri-Bojd S, Balali-
Mood M and Mousavi SH (2018). Investigating the influence of
polyplex size on toxicity properties of polyethylenimine mediated
gene delivery. Life Sci 197: 101-108.
Kunath K, von Harpe A, Fischer D, Petersen H, Bickel U and Voigt
K (2003). Low-molecular-weight polyethylenimine as a non-viral
vector for DNA delivery: comparison of physicochemical
properties, transfection efficiency and in vivo distribution with
high-molecular-weight polyethylenimine. J Control Release Off J
Control Release Soc 89(1): 113-125.
Lächelt U and Wagner E (2015). Nucleic Acid Therapeutics Using
Polyplexes: A Journey of 50 Years (and Beyond). Chem Rev
115(19): 11043-1078.
Lam AP and Dean DA (2010). Progress and prospects: nuclear
import of non-viral vectors. Gene Ther 17(4): 439-447.
Leclercq L, Reinhard S, Chamieh J, Döblinger M, Wagner E and
Cottet H (2015). Fast Characterization of Polyplexes by Taylor
Dispersion Analysis. Macromolecules 48(19): 7216-721.
Li S-D and Huang L (2006). Targeted Delivery of Antisense
Oligodeoxynucleotide and Small Interference RNA into Lung
Cancer Cells. Mol Pharm 3(5): 579-588.
Männistö M, Reinisalo M, Ruponen M, Honkakoski P, Tammi M and
Urtti A (2007). Polyplex-mediated gene transfer and cell cycle:
effect of carrier on cellular uptake and intracellular kinetics, and
significance of glycosaminoglycans. J Gene Med 9(6): 479-487.
Martinez-Fong D, Bannon MJ, Trudeau L-E, Gonzalez-Barrios JA,
Arango-Rodriguez ML and Hernandez-Chan NG (2012). NTS-
Polyplex: a potential nanocarrier for neurotrophic therapy of
Parkinson’s disease. Nanomedicine Nanotechnol Biol Med 8(7):
1052-1069.
Mockey M, Bourseau E, Chandrashekhar V, Chaudhuri A, Lafosse S
and Le Cam E (2007). mRNA-based cancer vaccine: prevention
of B16 melanoma progression and metastasis by systemic
injection of MART1 mRNA histidylated lipopolyplexes. Cancer
Gene Ther 14(9): 802-914.
Moghimi SM, Symonds P, Murray JC, Hunter AC, Debska G and
Szewczyk A (2005). A two-stage poly(ethylenimine)-mediated
cytotoxicity: implications for gene transfer/therapy. Mol Ther
11(6): 990-995.
Novo L, Mastrobattista E, van Nostrum CF, Lammers T and
Hennink WE (2015). Decationized polyplexes for gene delivery.
Expert Opin Drug Deliv 12(4): 507-512.
Ogris M, Steinlein P, Carotta S, Brunner S and Wagner E (2001).
DNA/polyethylenimine transfection particles: Influence of
ligands, polymer size, and PEGylation on internalization and
gene expression. AAPS Pharm Sci 3(3): 43-53.
Petersen H, Fechner PM, Martin AL, Kunath K, Stolnik S and
Roberts CJ (2002). Polyethylenimine-graft-poly (ethylene glycol)
4686 Int J Pharm Sci Nanotech Vol 12; Issue 6 • November− December 2019
copolymers: influence of copolymer block structure on DNA
complexation and biological activities as gene delivery system.
Bioconjug Chem 13(4): 845-854.
Ramamoorth M (2015). Non-Viral Vectors in Gene Therapy- An
Overview. J Clin Diagn Res 9(1): GE01-GE06.
Rejman J, Conese M and Hoekstra D (2006). Gene Transfer by
Means of Lipo- and Polyplexes: Role of Clathrin and Caveolae-
Mediated Endocytosis. J Liposome Res 16(3): 237-247.
Rubio-Zapata HA, Rembao-Bojorquez JD, Arango-Rodriguez ML,
Dupouy S, Forgez P and Martinez-Fong D (2009). NT-polyplex: a
new tool for therapeutic gene delivery to neuroblastoma tumors.
Cancer Gene Ther 16(7): 573-584.
Taranejoo S, Liu J, Verma P and Hourigan K (2015). A review of the
developments of characteristics of PEI derivatives for gene
delivery applications. J Appl Polym Sci 132(25).
Troiber C, Kasper JC, Milani S, Scheible M, Martin I and Schaubhut
F (2013). Comparison of four different particle sizing methods
for siRNA polyplex characterization. Eur J Pharm Biopharm
84(2): 255-264.
Tros de Ilarduya C, Sun Y and Düzgüneş N (2010). Gene delivery by
lipoplexes and polyplexes. Eur J Pharm Sci 40(3): 159-170.
Urbiola K, García L, Zalba S, Garrido MJ and Tros de Ilarduya C
(2013). Efficient serum-resistant lipopolyplexes targeted to the
folate receptor. Eur J Pharm Biopharm 83(3): 358-363.
van der Aa MAEM, Huth US, Häfele SY, Schubert R, Oosting RS
and Mastrobattista E (2007). Cellular Uptake of Cationic
Polymer-DNA Complexes Via Caveolae Plays a Pivotal Role in
Gene Transfection in COS-7 Cells. Pharm Res 24(8): 1590-1598.
Venkiteswaran S, Thomas T and Thomas TJ (2016). Selectivity of
polyethyleneimines on DNA nanoparticle preparation and gene
transport. Chemistry Select 1(6):1144-1150.
Von Gersdorff K, Sanders NN, Vandenbroucke R, De Smedt SC,
Wagner E and Ogris M (2006). The Internalization Route
Resulting in Successful Gene Expression Depends on both Cell
Line and Polyethylenimine Polyplex Type. Mol Ther 14(5): 745-
753.
Wagner E (2004). Strategies to Improve DNA Polyplexes for in Vivo
Gene Transfer: Will “Artificial Viruses” Be the Answer? Pharm
Res 21(1): 8-14.
Weide B, Garbe C, Rammensee H-G and Pascolo S (2008). Plasmid
DNA- and messenger RNA-based anti-cancer vaccination.
Immunol Lett 115(1): 33-42.
Address correspondence to: Rohan Aggarwal, B801 New Rajput partment
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Tel: +91-9717727487
E-mail: rohanaggarwal30@gmail.com