Formulations which increase the size of lipoplexes prevent serum-associated inhibition of transfection

Turek, Joseph W.; Dubertret, Catherine; Jaslin, Gabrielle; Antonakis, Kostas; Scherman, Daniel; Pitard, Bruno

doi:10.1002/(sici)1521-2254(200001/02)2:1<32::aid-jgm78>3.0.co;2-u

Cited by 102 publications

(75 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is reported that serum components such as negatively charged proteins cause the disintegration of their complex. [29][30][31][32][33][34][35] Our results support it, one of the reasons of less transfection activity of Lipofectamine TM 2000 in the presence of serum will be derived from the disintegration of liposome-dsRNA complex and resulting in the degradation of bound dsRNAs. DsRBD3 is seemed to retain the ability to interact with shRNA even in the circumstance of high serum concentration.…”

Section: Discussionsupporting

confidence: 81%

Improving the Stability of Short Hairpin RNA against Fetal Bovine Serum Using the Third Double-Stranded RNA-Binding Domain from Staufen Protein

Kawaguchi

Danjo

Okuda

et al. 2009

Biological & Pharmaceutical Bulletin

View full text Add to dashboard Cite

RNA interference (RNAi) is a technology that specifically inhibits gene expression and is carried out by small 21-27-nucleotide double-stranded small interfering RNA (siRNA) or short hairpin RNA (shRNA). RNAi has become a very promising technology for genetic research, however problems remain with the delivery of siRNA into cells. SiRNA and shRNA are easily degraded by RNases in body fluids and are hardly able to permeate cell membranes because of hydrophilic, polyanionic macromolecules which make their bioavailability very low. We have focused on the third double-stranded RNA-binding domain (dsRBD3) from the Mus musculus Staufen protein in order to develop a non-viral, multifunctional, artificial virus-like delivery system. We constructed a dsRBD3 expression vector and the recombinant dsRBD3 was expressed as a fusion protein with affinity tags. Purified dsRBD3 was mixed with siRNA or shRNA at various ratios and then added to fetal bovine serum (FBS) to evaluate the inhibition of the degradation of double-stranded RNA (dsRNA) by RNase. Unexpectedly, dsRBD3 was not able to protect the siRNA against degradation by FBS, but shRNA was stabilized to some degree by dsRBD3.

show abstract

Section: Discussionsupporting

confidence: 81%

Improving the Stability of Short Hairpin RNA against Fetal Bovine Serum Using the Third Double-Stranded RNA-Binding Domain from Staufen Protein

Kawaguchi

Danjo

Okuda

et al. 2009

Biological & Pharmaceutical Bulletin

View full text Add to dashboard Cite

show abstract

“…However, it has been reported that large-sized, colloidally unstable lipid/DNA complexes of over 700 nm diameter transfected more efficiently than colloidally stable 250 nm complexes. 29 It is yet to be determined what if any effect particle size of P[NIPAM/DMAEMA] and P[NIPAM/DMAEMAQ] polyplexes has on their DNA delivery potential in either in vitro or in vivo applications. P[NIPAM/DMAEMA] TCNGs of low DMAEMA content (8.3%mol) undergo extremely rapid dehydration and deswelling in water that results in loss of particle stability and aggregation.…”

Section: Discussionmentioning

confidence: 99%

Characterization of complexation of poly (N-isopropylacrylamide-co-2-(dimethylamino) ethyl methacrylate) thermoresponsive cationic nanogels with salmon sperm DNA

Moselhy

Vira²,

Liu³

et al. 2009

IJN

View full text Add to dashboard Cite

Thermoresponsive cationic nanogel (TCNG) networks based on N-isopropylacrylamide (NIPAM), 2-(dimethylamino)ethyl methacrylate (DMAEMA), and quaternary alkyl ammonium halide salts of DMAEMA (DMAEMAQ) were synthesized by dispersion polymerization technique. The thermoresponsive properties of TCNGs and TCNG-salmon sperm DNA (sasDNA) polyplexes were characterized in aqueous media of various pH and ionic strength. The distinct phase-transition properties of P[NIPAM/DMAEMA]/sasDNA and P[NIPAM/ DMAEMAQ]/sasDNA polyplexes were attributed to the condensing capability of polycations and to differences in the spatial distribution of structural charges in quaternized and nonquaternized networks. The findings demonstrate that stable TCNGs can be prepared with controllable responsive properties determined by the nature of the cationic charge incorporated and may have potential as vehicles for DNA delivery.

show abstract

“…[18][19][20] Increased transfection activity was observed when the NP-OH ternary complex was formed at concentrations of NaCl above 50 mM, even though the size was not significantly different with that formed in water (Figs. 5A, B).…”

Section: Discussionmentioning

confidence: 99%

Low-Molecular-Weight Polyethylenimine Enhanced Gene Transfer by Cationic Cholesterol-Based Nanoparticle Vector

Hattori

Maitani

2007

Biological & Pharmaceutical Bulletin

View full text Add to dashboard Cite

Both polyethylenimine (PEI) polymers and cationic nanoparticles have been widely used for non-viral DNA transfection. Previously, we reported that cationic nanoparticles composed of cholesteryl-3b b-carboxyamidoethylene-N-hydroxyethylamine and Tween 80 (NP-OH) could deliver plasmid DNA (pDNA) with high transfection efficiency. To increase the transfection activity of NP-OH, we investigated the potential synergism of PEI and NP-OH for the transfection of DNA into human prostate tumor PC-3, human cervices tumor Hela, and human lung adenocarcinoma A549 cells. The transfection efficiency with low-molecular PEI (MW 600) was low, but that with a combination of NP-OH and PEI was higher than with NP-OH alone, being comparable to commercially available lipofectamine 2000 and lipofectamine LTX, with very low cytotoxicity. Low-molecular weight PEI could not compact pDNA in size, but rather might help to dissociate pDNA from the complex and release pDNA from the endosome to cytoplasm by the proton sponge effect. Therefore, the combination of cationic cholesterol-based nanoparticles and a low-molecular PEI has potential as a non-viral DNA vector for gene delivery.

show abstract

Formulations which increase the size of lipoplexes prevent serum-associated inhibition of transfection

Cited by 102 publications

References 26 publications

Improving the Stability of Short Hairpin RNA against Fetal Bovine Serum Using the Third Double-Stranded RNA-Binding Domain from Staufen Protein

Improving the Stability of Short Hairpin RNA against Fetal Bovine Serum Using the Third Double-Stranded RNA-Binding Domain from Staufen Protein

Characterization of complexation of poly (N-isopropylacrylamide-co-2-(dimethylamino) ethyl methacrylate) thermoresponsive cationic nanogels with salmon sperm DNA

Low-Molecular-Weight Polyethylenimine Enhanced Gene Transfer by Cationic Cholesterol-Based Nanoparticle Vector

Contact Info

Product

Resources

About