Design of embedded chimeric peptide nucleic acids that efficiently enter and accurately reactivate gene expression in vivo.

Abstract:

:Pharmacological treatments designed to reactivate fetal γ-globin can lead to an effective and successful clinical outcome in patients with hemoglobinopathies. However, new approaches remain highly desired because such treatments are not equally effective for all patients, and toxicity issues remain. We have taken a systematic approach to develop an embedded chimeric peptide nucleic acid (PNA) that effectively enters the cell and the nucleus, binds to its target site at the human fetal γ-globin promoter, and reactivates this transcript in adult transgenic mouse bone marrow and human primary peripheral blood cells. In vitro and in vivo DNA-binding assays in conjunction with live-cell imaging have been used to establish and optimize chimeric PNA design parameters that lead to successful gene activation. Our final molecule contains a specific γ-promoter-binding PNA sequence embedded within two amino acid motifs: one leads to efficient cell/nuclear entry, and the other generates transcriptional reactivation of the target. These embedded PNAs overcome previous limitations and are generally applicable to the design of in vivo transcriptional activation reagents that can be directed to any promoter region of interest and are of direct relevance to clinical applications that would benefit from such a need.

authors

Chen J,Peterson KR,Iancu-Rubin C,Bieker JJ

doi

10.1073/pnas.0912896107

subject

Has Abstract

pub_date

2010-09-28 00:00:00

pages

16846-51

issue

39

eissn

0027-8424

issn

1091-6490

pii

0912896107

journal_volume

107

pub_type

杂志文章
  • The criminogenic and psychological effects of police stops on adolescent black and Latino boys.

    abstract::Proactive policing, the strategic targeting of people or places to prevent crimes, is a well-studied tactic that is ubiquitous in modern law enforcement. A 2017 National Academies of Sciences report reviewed existing literature, entrenched in deterrence theory, and found evidence that proactive policing strategies can...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1808976116

    authors: Del Toro J,Lloyd T,Buchanan KS,Robins SJ,Bencharit LZ,Smiedt MG,Reddy KS,Pouget ER,Kerrison EM,Goff PA

    更新日期:2019-04-23 00:00:00

  • Night-vision brain area in migratory songbirds.

    abstract::Twice each year, millions of night-migratory songbirds migrate thousands of kilometers. To find their way, they must process and integrate spatiotemporal information from a variety of cues including the Earth's magnetic field and the night-time starry sky. By using sensory-driven gene expression, we discovered that ni...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0409575102

    authors: Mouritsen H,Feenders G,Liedvogel M,Wada K,Jarvis ED

    更新日期:2005-06-07 00:00:00

  • Increasing aridity reduces soil microbial diversity and abundance in global drylands.

    abstract::Soil bacteria and fungi play key roles in the functioning of terrestrial ecosystems, yet our understanding of their responses to climate change lags significantly behind that of other organisms. This gap in our understanding is particularly true for drylands, which occupy ∼41% of Earth´s surface, because no global, sy...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1516684112

    authors: Maestre FT,Delgado-Baquerizo M,Jeffries TC,Eldridge DJ,Ochoa V,Gozalo B,Quero JL,García-Gómez M,Gallardo A,Ulrich W,Bowker MA,Arredondo T,Barraza-Zepeda C,Bran D,Florentino A,Gaitán J,Gutiérrez JR,Huber-Sannwald E,Jan

    更新日期:2015-12-22 00:00:00

  • Trans/cis (Z/E) photoisomerization of the chromophore of photoactive yellow protein is not a prerequisite for the initiation of the photocycle of this photoreceptor protein.

    abstract::The chromophore of photoactive yellow protein (PYP) (i.e., 4-hydroxycinnamic acid) has been replaced by an analogue with a triple bond, rather than a double bond (by using 4-hydroxyphenylpropiolic acid in the reconstitution, yielding hybrid I) and by a "locked" chromophore (through reconstitution with 7-hydroxycoumari...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.95.13.7396

    authors: Cordfunke R,Kort R,Pierik A,Gobets B,Koomen GJ,Verhoeven JW,Hellingwerf KJ

    更新日期:1998-06-23 00:00:00

  • Cloning of a human complement component C4 gene.

    abstract::Six overlapping cosmid clones having an average insert size of 40 kilobase pairs were identified and isolated from a human genomic library by using a cDNA probe, Alu-7, specific for the amino acid sequence of C4d, a known region of the fourth component of human complement. Analysis of these genomic clones by restricti...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.80.1.264

    authors: Carroll MC,Porter RR

    更新日期:1983-01-01 00:00:00

  • Cancer dormancy and cell signaling: induction of p21(waf1) initiated by membrane IgM engagement increases survival of B lymphoma cells.

    abstract::The p21(WAF1) (p21) cyclin-dependent kinase inhibitor plays a major role in regulating cell cycle arrest. It was recently reported that the p53-independent elevation of p21 protein levels is essential in mediating the G(1) arrest resulting from signal transduction events initiated by the crosslinking of membrane IgM o...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.96.15.8711

    authors: Marches R,Hsueh R,Uhr JW

    更新日期:1999-07-20 00:00:00

  • Remodeling of synaptic architecture during hippocampal "kindling".

    abstract::The "kindling" phenomenon is associated with long-lasting facilitation of synaptic transmission. A possible mechanism of such facilitation could involve changes in the number of synaptic contacts. However, previous attempts to demonstrate a synaptic morphological alteration that could account for the long-term effects...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.85.9.3260

    authors: Geinisman Y,Morrell F,deToledo-Morrell L

    更新日期:1988-05-01 00:00:00

  • Lambda YES: a multifunctional cDNA expression vector for the isolation of genes by complementation of yeast and Escherichia coli mutations.

    abstract::This work describes a multifunctional phage lambda expression vector system, lambda YES, designed to facilitate gene isolation from eukaryotes by complementation of Escherichia coli and Saccharomyces cerevisiae mutations. lambda YES vectors have a selection for cDNA inserts using an oligo adaptor strategy and are capa...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.88.5.1731

    authors: Elledge SJ,Mulligan JT,Ramer SW,Spottswood M,Davis RW

    更新日期:1991-03-01 00:00:00

  • Effect of selenium deficiency on cuticle integrity in the Cladocera (Crustacea).

    abstract::Daphnia pulex de Geer and Daphnia magna Straus populations cannot be maintained in defined (sensu stricto) media containing less than 0.1 part per billion (ppb) of selenium. A concentration of 1 ppb is sufficient to satisfy minimal needs in otherwise sufficient media. In the first generation with no selenium added to ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.81.11.3433

    authors: Keating KI,Dagbusan BC

    更新日期:1984-06-01 00:00:00

  • Unique DNA associated with mitochondria in the "S"-type cytoplasm of male-sterile maize.

    abstract::Mitochondrial DNAs were prepared from maize lines with normal cytoplasm and with the T, C, S, and EP sources of male-sterile cytoplasms. Agarose gel electrophoresis of these preparations revealed a main high-molecular-weight DNA band. In addition, the S cytoplasm was characterized by the presence of two faster migrati...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.74.7.2904

    authors: Pring DR,Levings CS,Hu WW,Timothy DH

    更新日期:1977-07-01 00:00:00

  • Failure of the opiate antagonist naloxone to modify hypnotic analgesia.

    abstract::Hypnotic analgesia in some respects resembles opiate analgesia. We tested the hypothesis that some features of hypnotic analgesia are mediated through neuronal pathways activating specific opiate receptors in brain. The opiate antagonist naloxone had no effect on hypnotic analgesia in three subjects. Thus, the hypothe...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.72.6.2041

    authors: Goldstein A,Hilgard ER

    更新日期:1975-06-01 00:00:00

  • On the transfer of integral proteins into membranes.

    abstract::We have earlier proposed a molecular mechanism for the translocation of hydrophilic proteins across membranes that accounts for the experimental facts and meets the restrictions that we stipulate for such a mechanism. In particular, the restrictions are that translocation occurs by successive segments of the polypepti...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.84.7.1960

    authors: Singer SJ,Maher PA,Yaffe MP

    更新日期:1987-04-01 00:00:00

  • The human Bloom syndrome gene suppresses the DNA replication and repair defects of yeast dna2 mutants.

    abstract::Bloom syndrome is a disorder of profound and early cancer predisposition in which cells become hypermutable, exhibit high frequency of sister chromatid exchanges, and show increased micronuclei. BLM, the gene mutated in Bloom syndrome, has been cloned previously, and the BLM protein is a member of the RecQ family of D...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1431624100

    authors: Imamura O,Campbell JL

    更新日期:2003-07-08 00:00:00

  • In vivo aminoacylation of human and Xenopus suppressor tRNAs constructed by site-specific mutagenesis.

    abstract::Amber suppressor tRNA genes were constructed by site-specific mutagenesis of the anticodons of human lysine-inserting tRNA (tRNA(Lys)) and glutamine-inserting tRNA (tRNA(Gln)) genes, and a Xenopus laevis tyrosine-inserting tRNA (tRNA(Tyr)) gene. As previous in vitro studies in prokaryotes have shown that substitution ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.84.8.2185

    authors: Ho YS,Kan YW

    更新日期:1987-04-01 00:00:00

  • Synthesis of the next-generation therapeutic antibodies that combine cell targeting and antibody-catalyzed prodrug activation.

    abstract::An obstacle in the utilization of catalytic Abs for selective prodrug activation in cancer therapy has been systemic tumor targeting. Here we report the generation of catalytic Abs that effectively target tumor cells with undiminished prodrug activation capability. Ab conjugates were prepared by covalent conjugation o...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0700223104

    authors: Abraham S,Guo F,Li LS,Rader C,Liu C,Barbas CF 3rd,Lerner RA,Sinha SC

    更新日期:2007-03-27 00:00:00

  • Primary structure of Escherichia coli ribosomal protein S1 and of its gene rpsA.

    abstract::The primary structure of proteins S1, the largest protein component of the Escherichia coli ribosome, has been elucidated by determining the amino acid sequence of the protein (from E. coli MRE600) and the nucleotide sequence of the S1 gene (rpsA, of a K-12 strain). The two methods gave results in perfect agreement ex...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.79.4.1008

    authors: Schnier J,Kimura M,Foulaki K,Subramanian AR,Isono K,Wittmann-Liebold B

    更新日期:1982-02-01 00:00:00

  • Ig gene-like molecule CD31 plays a nonredundant role in the regulation of T-cell immunity and tolerance.

    abstract::CD31 is an Ig-like molecule expressed by leukocytes and endothelial cells with an established role in the regulation of leukocyte trafficking. Despite genetic deletion of CD31 being associated with exacerbation of T cell-mediated autoimmunity, the contribution of this molecule to T-cell responses is largely unknown. H...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1011748107

    authors: Ma L,Mauro C,Cornish GH,Chai JG,Coe D,Fu H,Patton D,Okkenhaug K,Franzoso G,Dyson J,Nourshargh S,Marelli-Berg FM

    更新日期:2010-11-09 00:00:00

  • The final acylation step in taxol biosynthesis: cloning of the taxoid C13-side-chain N-benzoyltransferase from Taxus.

    abstract::The formation of several acyl groups and an amide group of Taxol is catalyzed by regioselective CoA thioester-dependent acyltransferases. Several full-length acyltransferase sequences, obtained from a cDNA library constructed from mRNA isolated from Taxus cuspidata cells induced for Taxol production with methyl jasmon...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.082115799

    authors: Walker K,Long R,Croteau R

    更新日期:2002-07-09 00:00:00

  • The surface oxidation potential of human neuromelanin reveals a spherical architecture with a pheomelanin core and a eumelanin surface.

    abstract::Neuromelanin (NM) isolated from the substantia nigra region of the human brain was studied by scanning probe and photoelectron emission microscopies. Atomic force microscopy reveals that NM granules are comprised of spherical structures with a diameter of approximately 30 nm, similar to that observed for Sepia cuttlef...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0604010103

    authors: Bush WD,Garguilo J,Zucca FA,Albertini A,Zecca L,Edwards GS,Nemanich RJ,Simon JD

    更新日期:2006-10-03 00:00:00

  • Isolation of an ovine pulmonary surfactant-associated anionic peptide bactericidal for Pasteurella haemolytica.

    abstract::Ovine pulmonary surfactant is bactericidal for Pasteurella haemolytica when surfactant and bacteria mixtures are incubated with normal ovine serum. To isolate this component, surfactant (1 mg/ml) was centrifuged at 100,000 x gav, and the supernatant was fractionated by HPLC. Fractions were eluted with acetonitrile (10...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.93.1.412

    authors: Brogden KA,De Lucca AJ,Bland J,Elliott S

    更新日期:1996-01-09 00:00:00

  • Natal dispersal driven by environmental conditions interacting across the annual cycle of a migratory songbird.

    abstract::Natal dispersal, the process through which immature individuals permanently depart their natal area in search of new sites, is integral to the ecology and evolution of animals. Insights about the underlying causes of natal dispersal arise mainly from research on species whose short dispersal distances or restricted di...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0710732105

    authors: Studds CE,Kyser TK,Marra PP

    更新日期:2008-02-26 00:00:00

  • Gene therapy of murine teratocarcinoma: separate functions for insulin-like growth factors I and II in immunogenicity and differentiation.

    abstract::Teratocarcinoma is a germ-line carcinoma giving rise to an embryoid tumor with structures derived from the three embryonic layers: mesoderm, endoderm, and ectoderm. Teratocarcinoma is widely used as an in vitro model system to study regulation of cell determination and differentiation during mammalian embryogenesis. M...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.91.13.6088

    authors: Trojan J,Johnson TR,Rudin SD,Blossey BK,Kelley KM,Shevelev A,Abdul-Karim FW,Anthony DD,Tykocinski ML,Ilan J

    更新日期:1994-06-21 00:00:00

  • Uncoupling reproduction from metabolism extends chronological lifespan in yeast.

    abstract::Studies of replicative and chronological lifespan in Saccharomyces cerevisiae have advanced understanding of longevity in all eukaryotes. Chronological lifespan in this species is defined as the age-dependent viability of nondividing cells. To date this parameter has only been estimated under calorie restriction, mimi...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1323918111

    authors: Nagarajan S,Kruckeberg AL,Schmidt KH,Kroll E,Hamilton M,McInnerney K,Summers R,Taylor T,Rosenzweig F

    更新日期:2014-04-15 00:00:00

  • Contraction of detergent-treated smooth muscle.

    abstract::After exposure of segments of rabbit taenia coli to the nonionic detergent Triton X-100, tension could be induced by increasing the [Ca2+] in the micromolar range. In the presence of a saturating [Ca2+], this preparation developed nearly 100% of the control tetanus tension recorded from the intact muscle prior to the ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.75.7.3527

    authors: Gordon AR

    更新日期:1978-07-01 00:00:00

  • Cytoplasmic transfer of resistance to antimycin A in Chinese hamster cells.

    abstract::A mutant subline of V79 Chinese hamster cells resistant to antimycin A (ANT) was obtained by treatment with ethyl methanesulfonate followed by serial selection in ANT-containing medium. Clonal derivatives of the resistant line are less susceptible than parent cells to growth inhibition by ANT in mass populations and s...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.75.11.5604

    authors: Harris M

    更新日期:1978-11-01 00:00:00

  • Global synthesis of conservation studies reveals the importance of small habitat patches for biodiversity.

    abstract::Island biogeography theory posits that species richness increases with island size and decreases with isolation. This logic underpins much conservation policy and regulation, with preference given to conserving large, highly connected areas, and relative ambivalence shown toward protecting small, isolated habitat patc...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.1813051115

    authors: Wintle BA,Kujala H,Whitehead A,Cameron A,Veloz S,Kukkala A,Moilanen A,Gordon A,Lentini PE,Cadenhead NCR,Bekessy SA

    更新日期:2019-01-15 00:00:00

  • Requirement of Nanog dimerization for stem cell self-renewal and pluripotency.

    abstract::Pluripotency of embryonic stem (ES) cells is maintained by transcription factors that form a highly interconnected protein interaction network surrounding the homeobox protein Nanog. Enforced expression of Nanog in mouse ES (mES) cells promotes self-renewal and alleviates their requirement for leukemia inhibitory fact...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.0802288105

    authors: Wang J,Levasseur DN,Orkin SH

    更新日期:2008-04-29 00:00:00

  • Specific binding of erythropoietin to its receptor on responsive mouse erythroleukemia cells.

    abstract::Erythropoietin (Epo) is a glycoprotein factor that specifically regulates the proliferation and differentiation of erythroid progenitor cells. Here we describe the isolation of Epo-responsive mouse erythroleukemia cell line SKT6, the characterization of the specific binding of biologically active 125I-labeled human Ep...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.84.12.4126

    authors: Todokoro K,Kanazawa S,Amanuma H,Ikawa Y

    更新日期:1987-06-01 00:00:00

  • Genetic analysis of microtubule motor proteins in Drosophila: a mutation at the ncd locus is a dominant enhancer of nod.

    abstract::The nod (no distributive disjunction) and the ncd (non-claret disjunctional) mutations are both female-specific, recessive meiotic mutations in Drosophila melanogaster. Mutations at either locus show high frequencies of nondisjunction at meiosis I and both have been shown to encode kinesin-like proteins. Unlike the nc...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.88.16.7165

    authors: Knowles BA,Hawley RS

    更新日期:1991-08-15 00:00:00

  • Probing the open state of cytochrome P450cam with ruthenium-linker substrates.

    abstract::Cytochromes P450 play key roles in drug metabolism and disease by oxidizing a wide variety of natural and xenobiotic compounds. High-resolution crystal structures of P450cam bound to ruthenium sensitizer-linked substrates reveal an open conformation of the enzyme that allows substrates to access the active center via ...

    journal_title:Proceedings of the National Academy of Sciences of the United States of America

    pub_type: 杂志文章

    doi:10.1073/pnas.221297998

    authors: Dunn AR,Dmochowski IJ,Bilwes AM,Gray HB,Crane BR

    更新日期:2001-10-23 00:00:00