Artificial DNA: PNA & XNA最新文献_第5页

Cell number and transfection volume dependent peptide nucleic acid antisense activity by cationic delivery methods. 细胞数量和转染体积依赖于阳离子递送方法的肽核酸反义活性。

Artificial DNA: PNA & XNA Pub Date : 2012-01-01 DOI: 10.4161/adna.19906

Laia Llovera, Peter Berthold, Peter E Nielsen, Takehiko Shiraishi

{"title":"Cell number and transfection volume dependent peptide nucleic acid antisense activity by cationic delivery methods.","authors":"Laia Llovera, Peter Berthold, Peter E Nielsen, Takehiko Shiraishi","doi":"10.4161/adna.19906","DOIUrl":"https://doi.org/10.4161/adna.19906","url":null,"abstract":"Efficient intracellular delivery is essential for high activity of nucleic acids based therapeutics, including antisense agents. Several strategies have been developed and practically all rely on auxiliary transfection reagents such as cationic lipids, cationic polymers and cell penetrating peptides as complexing agents and carriers of the nucleic acids. However, uptake mechanisms remain rather poorly understood, and protocols always require optimization of transfection parameters. Considering that cationic transfection complexes bind to and thus may up-concentrate on the cell surface, we have now quantitatively compared the cellular activity (in the pLuc705 HeLa cell splice correction system) of PNA antisense oligomers using lipoplex delivery of cholesterol- and bisphosphonate-PNA conjugates, polyplex delivery via a PNA-polyethyleneimine conjugate and CPP delivery via a PNA-octaarginine conjugate upon varying the cell culture transfection volume (and cell density) at fixed PNA concentration. The results show that for all delivery modalities the cellular antisense activity increases (less than proportionally) with increasing volume (in some cases accompanied with increased toxicity), and that this effect is more pronounced at higher cell densities. These results emphasize that transfection efficacy using cationic carriers is critically dependent on parameters such as transfection volume and cell density, and that these must be taken into account when comparing different delivery regimes.","PeriodicalId":8444,"journal":{"name":"Artificial DNA: PNA & XNA","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2012-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4161/adna.19906","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"30675681","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 9

(α,α-dimethyl)glycyl (dmg) PNAs: achiral PNA analogs that form stronger hybrids with cDNA relative to isosequential RNA. (α，α-二甲基)甘酰基(dmg) PNAs:相对于同序RNA，与cDNA形成更强杂交的非手性PNA类似物。

Artificial DNA: PNA & XNA Pub Date : 2012-01-01 DOI: 10.4161/adna.19185

Aland Gourishankar, Krishna N Ganesh

{"title":"(α,α-dimethyl)glycyl (dmg) PNAs: achiral PNA analogs that form stronger hybrids with cDNA relative to isosequential RNA.","authors":"Aland Gourishankar, Krishna N Ganesh","doi":"10.4161/adna.19185","DOIUrl":"https://doi.org/10.4161/adna.19185","url":null,"abstract":"The design and facile synthesis of sterically constrained new analogs of PNA having gem-dimethyl substitutions on glycine (dmg-PNA-T) is presented. The PNA oligomers [aminoethyl dimethylglycyl (aedmg) and aminopropyl dimethylglycyl (apdmg)] synthesized from the monomers 6 and 12) effected remarkable stabilization of homothyminePNA(2):homoadenine DNA/RNA triplexes and mixed base sequence duplexes with target cDNA or RNA. They show a higher binding to DNA relative to that with isosequential RNA. This may be a structural consequence of the sterically rigid gem-dimethyl group, imposing a pre-organized conformation favorable for complex formation with cDNA. The results complement our previous work that had demonstrated that cyclohexanyl-PNAs favor binding with cRNA compared with cDNA and imply that the biophysical and structural properties of PNAs can be directed by introduction of the right rigidity in PNA backbone devoid of chirality. This approach of tweaking selectivity in binding of PNA constructs by installing gem-dimethyl substitution in PNA backbone can be extended to further fine-tuning by similar substitution in the aminoethyl segment as well either individually or in conjunction with present substitution.","PeriodicalId":8444,"journal":{"name":"Artificial DNA: PNA & XNA","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2012-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4161/adna.19185","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"30677913","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 9

Potent and sustained cellular inhibition of miR-122 by lysine-derivatized peptide nucleic acids (PNA) and phosphorothioate locked nucleic acid (LNA)/2'-O-methyl (OMe) mixmer anti-miRs in the absence of transfection agents. 赖氨酸衍生肽核酸(PNA)和磷酸化锁定核酸(LNA)/2'- o -甲基(OMe)混合抗mir在没有转染剂的情况下有效和持续的细胞抑制miR-122。

Artificial DNA: PNA & XNA Pub Date : 2011-07-01 DOI: 10.4161/adna.17731

Adrian G Torres, Richard N Threlfall, Michael J Gait

{"title":"Potent and sustained cellular inhibition of miR-122 by lysine-derivatized peptide nucleic acids (PNA) and phosphorothioate locked nucleic acid (LNA)/2'-O-methyl (OMe) mixmer anti-miRs in the absence of transfection agents.","authors":"Adrian G Torres, Richard N Threlfall, Michael J Gait","doi":"10.4161/adna.17731","DOIUrl":"https://doi.org/10.4161/adna.17731","url":null,"abstract":"Efficient cell delivery of antisense oligonucleotides (ONs) is a key issue for their potential therapeutic use. It has been shown recently that some ONs can be delivered into cells without the use of transfection agents (gymnosis), but this generally requires cell incubation over several days and high amounts of ONs (micromolar concentrations). Here we have targeted microRNA 122 (miR-122), a small non-coding RNA involved in regulation of lipid metabolism and in the replication of hepatitis C virus, with ONs of different chemistries (anti-miRs) by gymnotic delivery in cell culture. Using a sensitive dual-luciferase reporter assay, anti-miRs were screened for their ability to enter liver cells gymnotically and inhibit miR-122 activity. Efficient miR-122 inhibition was obtained with cationic PNAs and 2'-O-methyl (OMe) and Locked Nucleic Acids (LNA)/OMe mixmers containing either phosphodiester (PO) or phosphorothioate (PS) linkages at sub-micromolar concentrations when incubated with cells for just 4 hours. Furthermore, PNA and PS-containing anti-miRs were able to sustain miR-122 inhibitory effects for at least 4 days. LNA/OMe PS anti-miRs were the most potent anti-miR chemistry tested in this study, an ON chemistry that has been little exploited so far as anti-miR agents towards therapeutics.","PeriodicalId":8444,"journal":{"name":"Artificial DNA: PNA & XNA","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2011-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4161/adna.17731","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"30603156","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 26

PNA HyBeacons for analysis of human mutations related to statin-induced myopathy. PNA HyBeacons用于分析与他汀类药物诱导的肌病相关的人类突变。

Artificial DNA: PNA & XNA Pub Date : 2011-07-01 DOI: 10.4161/adna.18179

Nittaya Gale, Petr Kocalka, Charlotte Mardle, Tom Brown

引用次数: 0

Peptide nucleic acid (PNA) cell penetrating peptide (CPP) conjugates as carriers for cellular delivery of antisense oligomers. 肽核酸(PNA)细胞穿透肽(CPP)偶联物作为反义寡聚物的细胞递送载体。

Artificial DNA: PNA & XNA Pub Date : 2011-07-01 DOI: 10.4161/adna.18739

Takehiko Shiraishi, Peter E Nielsen

{"title":"Peptide nucleic acid (PNA) cell penetrating peptide (CPP) conjugates as carriers for cellular delivery of antisense oligomers.","authors":"Takehiko Shiraishi, Peter E Nielsen","doi":"10.4161/adna.18739","DOIUrl":"https://doi.org/10.4161/adna.18739","url":null,"abstract":"We have explored the merits of a novel delivery strategy for the antisense oligomers based on cell penetrating peptide (CPP) conjugated to a carrier PNA with sequence complementary to part of the antisense oligomer. The effect of these carrier CPP-PNAs was evaluated by using antisense PNA targeting splicing correction of the mutated luciferase gene in the HeLa pLuc705 cell line, reporting cellular (nuclear) uptake of the antisense PNA via luciferase activity measurement. Carrier CPP-PNA constructs were studied in terms of construct modification (with octaarginine and/or decanoic acid) and carrier PNA length (to adjust binding affinity). In general, the carrier CPP-PNA constructs including the ones with decanoyl modification provided significant increase of the activity of unmodified antisense PNA as well as of antisense octaarginine-PNA conjugates. Antisense activity, and by inference cellular delivery, of unmodified antisense PNA was enhanced at least 20-fold at 6 μM upon the complexation with an equimolar amount of nonamer carrier decanoyl-CPP-PNA (Deca-cPNA1(9)-(D-Arg)8). The antisense activity of a CPP-PNA ((D-Arg)8-asPNA) (at 2 μM) was improved 6-fold and 8-fold by a heptamer carrier CPP-PNA (cPNA1(7)-(D-Arg)8) and hexamer carrier decanoyl-CPP-PNA (Deca-cPNA1(6)-(D-Arg)8), respectively, without showing significant additional cellular toxicity. Most interestingly, the activity reached the same level obtained by enhancement with endosomolytic chloroquine (CQ) treatment, suggesting that the carrier might facilitate endosomal escape. Furthermore, 50% downregulation of luciferase expression at 60 nM siRNA was obtained using this carrier CPP-PNA delivery strategy (with CQ co-treatment) for a single stranded antisense RNA targeting normal luciferase mRNA. These results indicated that CPP-PNA carriers may be used as effective cellular delivery vectors for different types of antisense oligomers and also allows use of combinations of (at least two) different CPP ligands.","PeriodicalId":8444,"journal":{"name":"Artificial DNA: PNA & XNA","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2011-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4161/adna.18739","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"30603161","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 47

RNA-DNA sequence differences spell genetic code ambiguities. RNA-DNA序列的差异导致了遗传密码的模糊性。

Artificial DNA: PNA & XNA Pub Date : 2011-07-01 DOI: 10.4161/adna.17086

Thomas Bentin, Michael L Nielsen

引用次数: 1

Artificial DNA structures. 人工DNA结构。

Artificial DNA: PNA & XNA Pub Date : 2011-04-01 DOI: 10.4161/adna.2.2.17085

Peter E Nielsen

引用次数: 0

Noncovalent binding and fluorogenic response of cyanine dyes to DNA homoquadruplex and PNA-DNA heteroquadruplex structures. 花青素染料对DNA同型四重体和PNA-DNA异型四重体结构的非共价结合和荧光反应。

Artificial DNA: PNA & XNA Pub Date : 2011-04-01 DOI: 10.4161/adna.2.2.16339

Halimatu S Mohammed, Junriz O Delos Santos, Bruce A Armitage

引用次数: 17

A ribozyme transcribed by a ribozyme. 核酶由核酶转录的核酶

Artificial DNA: PNA & XNA Pub Date : 2011-04-01 DOI: 10.4161/adna.2.2.16852

Thomas Bentin

引用次数: 1

Pyrrolidinyl peptide nucleic acid with α/β-peptide backbone: A conformationally constrained PNA with unusual hybridization properties. 具有α/β-肽骨架的吡咯烷基肽核酸:一种具有不同寻常杂交特性的构象约束的PNA。

Artificial DNA: PNA & XNA Pub Date : 2011-04-01 DOI: 10.4161/adna.2.2.16340

Chotima Vilaivan, Choladda Srisuwannaket, Cheeraporn Ananthanawat, Chaturong Suparpprom, Junji Kawakami, Yoshie Yamaguchi, Yuko Tanaka, Tirayut Vilaivan

{"title":"Pyrrolidinyl peptide nucleic acid with α/β-peptide backbone: A conformationally constrained PNA with unusual hybridization properties.","authors":"Chotima Vilaivan, Choladda Srisuwannaket, Cheeraporn Ananthanawat, Chaturong Suparpprom, Junji Kawakami, Yoshie Yamaguchi, Yuko Tanaka, Tirayut Vilaivan","doi":"10.4161/adna.2.2.16340","DOIUrl":"https://doi.org/10.4161/adna.2.2.16340","url":null,"abstract":"We describe herein a new conformationally constrained analog of PNA carrying an alternating α/β amino acid backbone consisting of (2'R,4'R)-nucleobase-subtituted proline and (1S,2S)-2-aminocyclopentanecarboxylic acid (acpcPNA). The acpcPNA has been synthesized and evaluated for DNA, RNA and self-pairing properties by thermal denaturation experiments. It can form antiparallel hybrids with complementary DNA with high affinity and sequence specificity. Unlike other PNA systems, the thermal stability of acpcPNA·DNA hybrid is largely independent of G+C contents, and is generally higher than that of acpcPNA·RNA hybrid with the same sequence. Thermodynamic parameters analysis suggest that the A·T base pairs in the acpcPNA·DNA hybrids are enthalpically stabilized over G·C pairs. The acpcPNA also shows a hitherto unreported behavior, namely the inability to form self-pairing hybrids. These unusual properties should make the new acpcPNA a potentially useful candidate for various applications including microarray probes and antigene agents.","PeriodicalId":8444,"journal":{"name":"Artificial DNA: PNA & XNA","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2011-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4161/adna.2.2.16340","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"29991668","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 35