RNA interference and its therapeutic potential

Ghosal, Anubrata; Kabir, Ahmad Humayan; Mandal, Abul

doi:10.2478/s11536-011-0005-5

Cited by 2 publications

(2 citation statements)

References 80 publications

(84 reference statements)

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“…siRNA continues to gain popularity in functional genomics studies and as a potential therapeutic that can target many diseases ranging from age-related macular degeneration to different types of cancer and viral infections. − Due to the highly negatively charged nature of the siRNA phosphate backbone, and due to its susceptibility to degradation by exo- and endonucleases, a carrier (vector) is required to ensure successful intracellular siRNA delivery to target cells. , Since being introduced as nonviral gene delivery vectors in 1987 by Felgner and co-workers, cationic lipids have been widely studied as nonviral gene and siRNA vectors. − …”

Section: Introductionmentioning

confidence: 99%

Efficient Silencing of EGFP Reporter Gene with siRNA Delivered by Asymmetrical N⁴,N⁹-Diacyl Spermines

et al. 2012

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It is important to obtain structure-activity relationship (SAR) data across cationic lipids for the self-assembly and nonviral intracellular delivery of siRNA. The aims of this work are to carry out a SAR study on the efficiency of asymmetrical N(4),N(9)-diacyl spermines in siRNA delivery and EGFP reporter gene silencing, with comparisons to selected mixtures composed of symmetrical N(4),N(9)-diacyl spermines. Another important aim of these studies is to quantify the changes in cell viability, assayed with alamarBlue, as a function of lipid structure. Therefore, we have designed, synthesized, purified, and assayed novel cationic lipids that are asymmetrical lipopolyamines based on spermine. Flow cytometry and fluorescence microscopy in an EGFP stably transfected HeLa cell line, measuring both delivery of fluorescently tagged siRNAs and silencing the EGFP signal, allowed quantitation of the differences between asymmetrical cationic lipids, mixtures of their symmetrical counterparts, and comparison with commercial nonviral delivery agents. Intracellular delivery of siRNA and gene silencing by siRNA differ with different hydrophobic domains. In these asymmetrical N(4),N(9)-diacyl spermines, lipids that enhance siRNA uptake do not necessarily enhance siRNA-induced inhibition of gene expression: C18 and longer saturated chains promote uptake, while more unsaturated C18 chains promote gene silencing. These properties are efficiently demonstrated in a new nontoxic cationic lipid siRNA vector, N(4)-linoleoyl-N(9)-oleoyl-1,12-diamino-4,9-diazadodecane (LinOS), which is also shown to be comparable with or superior to TransIT-TKO and Lipofectamine 2000.

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Section: Introductionmentioning

confidence: 99%

Efficient Silencing of EGFP Reporter Gene with siRNA Delivered by Asymmetrical N⁴,N⁹-Diacyl Spermines

et al. 2012

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“…RNAi based therapies emerged in the period following its discovery in 1998 in plants, and are promising therapeutic candidates to treat various types of diseases, ranging from age related macular oedema to respiratory tract infections to various types of cancer [34][35][36]. In addition to siRNA based therapies, shRNA [37,38] and miRNA [39] are potential therapeutic tools.…”

Section: Therapeutic Potential Of Rnai Based Therapiesmentioning

confidence: 99%

siRNA and Gene Formulation for Efficient Gene Therapy

Blagbrough¹,

Metwally²

2013

Gene Therapy - Tools and Potential Applications

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Gene Therapy -Tools and Potential Applications 136 plex. This core complex which cζrries-out mRN" degrζdζtion is the RN" induced silencing complex RISC [ -]. The degrζdζtion process requires the key ζrgonζute fζmily of proteins, which contζin ζ domζin with RNζse H endonucleζse type of ζctivity thζt cζtζlyse cleζvζge of the phosphodiester bonds of the tζrgeted mRN". RISC ζssembly ζnd subsequently its function to mediζte sequence specific mRN" degrζdζtion occur in the cytoplζsm of the cell [ ]. The source of the dsRN" segments incorporζted in RISC cζn be endogenously processed microRN" miRN" , short hζirpin RN" shRN" , or synthetic siRN". miRN" is produced from endogenous DN" through the ζction of RN" polymerζse II resulting in the formζtion of non-coding RN" cζlled primζry miRN" pri-miR-N" , which is processed in the nucleus by ζ protein complex contζining ζn enzyme known ζs Droshζ ζnd ζ dsRN" binding protein cofζctor cζlled Pζshζ DGCR . Droshζ cleζves pri-miRN" to produce pre-miRN" , ζ dsRN" of -nucleotides ζnd hζving ζ hζirpin loop, which binds to Exportin protein ζnd is trζnsferred from the nucleus into the cytoplζsm. Pre-miRN" is processed by Dicer RNζse III enzyme in the cytoplζsm to give miRN", typicζlly of nucleotides in length ζnd hζving two nucleotide overhζngs ζt the '-position [ , ], shRN" is produced by trζnscription from ζn exogenous DN" thζt is delivered to the nucleus, ζnd codes for ζ hζirpin shζped RN" with segments of length -nucleotides ζnd loop of nucleotides [ , ] which cζn then be processed by Dicer ζnd incorporζted in the RN"i mζchinery.Once in the cytoplζsm, the processed dsRN" miRN", processed shRN", or siRN" is then incorporζted into ζ protein complex RISC-loζding complex, RLC . In Drosophila the RLC is composed of the dsRN", heterodimer protein DCR Dicer vζriζnt /R D , possibly including the cζtζlytic ζrgonζute proteins ζs well in this complex. The ζctive RISC is formed when one of the RN" strζnds in the complex is cleζved the pζssenger strζnd ζnd the strζnd with the less thermodynζmic stζble '-end guide/ζnti-sense strζnd remζins in the complex. The mRN" with complementζry sequence to the guide strζnd binds to the ζctive RISC ζnd is cleζved by the endoribonucleζse ζctivity of the ζrgonζute component of the complex Figure . . . RNA duplex structure RN" is ζ polymer of ribonucleotides. Eζch RN" nucleotide is composed of one nucleobζse, the monosζcchζride pentose ribose, ζnd one phosphζte group. The nucleobζses in RN" ζre ζdenine purine bζse , guζnine purine bζse , urζcil pyrimidine bζse , ζnd cytosine pyrimidine bζse Figure . " nucleoside is formed when eζch bζse is connected viζ ζ glycosidic bond to the ζnomeric cζrbon ' of ribose, thus when glycosylζted, ζdenine, guζnine, urζcil, ζnd cytosine nucleobζses give ζdenosine, guζnosine, uridine, ζnd cytidine nucleosides. Eζch two nucleosides ζre connected viζ ζ phosphζte diester bond between the ' of one nucleoside ζnd ' of the next nucleoside to form the RN" polynucleotide strζnd. The mζin differences in the primζry structure of RN" ζnd DN" ...

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RNA interference and its therapeutic potential

Cited by 2 publications

References 80 publications

Efficient Silencing of EGFP Reporter Gene with siRNA Delivered by Asymmetrical N⁴,N⁹-Diacyl Spermines

Efficient Silencing of EGFP Reporter Gene with siRNA Delivered by Asymmetrical N⁴,N⁹-Diacyl Spermines

siRNA and Gene Formulation for Efficient Gene Therapy

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RNA interference and its therapeutic potential

Cited by 2 publications

References 80 publications

Efficient Silencing of EGFP Reporter Gene with siRNA Delivered by Asymmetrical N4,N9-Diacyl Spermines

Efficient Silencing of EGFP Reporter Gene with siRNA Delivered by Asymmetrical N4,N9-Diacyl Spermines

siRNA and Gene Formulation for Efficient Gene Therapy

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Efficient Silencing of EGFP Reporter Gene with siRNA Delivered by Asymmetrical N⁴,N⁹-Diacyl Spermines

Efficient Silencing of EGFP Reporter Gene with siRNA Delivered by Asymmetrical N⁴,N⁹-Diacyl Spermines