Nuclear Localization peptides - Creative Peptides

Cellular Import Mediated by Nuclear Localization Signal Peptide Sequences
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SV40 T-Ag-derived NLS peptide - GenScript

AB - Imaging the expression and localization of RNAs in live-cell nucleus can provide important information on RNA synthesis, processing, and transport. Here, we report the development of a bifunctional molecular beacon (NLS-MB) composed of a single nuclear localization sequence (NLS) peptide conjugated to a molecular beacon for efficient delivery and imaging of endogenous RNAs in the nuclei of living cells. We characterized the NLS-MBs by comparing their signal-to-noise ratios with unmodified molecular beacons and determined their efficiency of nuclear import. We demonstrated the specificity and sensitivity of the method by observing in living cells the localization and colocalization of small nuclear RNAs (snRNA) U1 and U2 at discrete foci in the nucleoplasm, and the localization of small nucleolar RNA U3 in the nucleolus. These snRNAs were chosen because of their essential roles in RNA biogenesis. The results were validated using in situ hybridization as positive control and random beacons as negative control. This novel approach may be applied to imaging other nuclear RNAs and pre-mRNAs in living cells.

03/01/2016 · Design and synthesis of NLS peptide-PEG-PA(G1.0)dendrimer-acridine conjugates
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Unit price ; 1 mg: SP-5244-1: 140 EUR : BUY ..

All reagents were the highest grade available. All used solvents were of HPLC grade and stored over molecular sieves 4A if necessary. Acridine 9-isothiocyanate was purchased from Acros Organic Co. PA(G1.0) dendrimer was obtained from Aldrich Chemical Co. Maleimide-PEG-carboxylate (M.w. 1000, 3400 and 5000) N-hydroxysuccinimide esters (MAL-PEG-NHS) were purchased from Nectar Pharmaceuticals Co. ExGen 500 was purchased from Fermentas Co. and Lipofectamine – from Invitrogen. NAP-10 and 25 columns were from GE Healthcare Co. NLS peptide with an additional cystein (CAPKKKRKVA-amide) was custom synthesized by Biomer Technology Co. and purified by the reverse phase HPLC.

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Nuclear translocation of plasmid DNA from cytoplasm is a bottleneck for efficient nonviral transfection. Various nuclear proteins are delivered into the nuclei by highly efficient signal-specific intracellular nuclear carriers. Short nuclear localization signal (NLS) sequences were found in most nuclear proteins. Multiple attempts of modifying nanocarriers or directly plasmid DNA with NLS peptides have been made to increase the efficacy of transfection, although with mostly controversial results-. The importance of “naked” plasmid DNA directly bound to NLS peptide is clear from the size of nuclear pore, not exceeding 30 nm in diameter. An intercalator, reversibly binding with DNA, would probably be the best means to directly anchor the NLS peptide; however, several factors should be taken in account to optimize the approach, such as number of peptides per DNA, accessibility of peptides for binding with nuclear carriers and efficiency of DNA protection in cytoplasm. In this paper we report the synthesis of novel DNA intercalating conjugates of PEG with linear NLS peptide and significant amelioration of DNA protection and results of lipofection and polyfection in vitro.

10/09/2008 · Six Classes of Nuclear Localization Signals Specific to Different ..
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of which one bears an NLS peptide, ..

The synthesis of the NLS-peptide intercalating arm is depicted in and involves three major steps: (1) conjugation of PEG with dendrimer, (2) reaction of dendrimer with acridine reagent, and (3) attachment of NLS-peptide to the distal end of PEG. Accessibility of NLS peptide to nuclear carriers is probably one of the crucial factors affecting nuclear translocation of DNA. Three different conjugates were obtained having PEG arm lengths of 1000, 3400 and 5000 Da. PA(G1.0)dendrimer was selected on the basis of molecular modeling and distances between planar base pairs in DNA duplex. Amount of intercalating moieties (acridines) has to be optimized, since, some preliminary data, suggested that single or paired intercalators may not be sufficient to ensure stable complex of the short arm-linked NLS peptide. Even more strong effect of PEG arm in our case forced us to choose tripled intercalators. Higher number of intercalators would form more stable complexes; however, slow dissociation of too strong complexes may reduce replication activity of the transfected DNA.

[5-FAM]-MPG∆NLS (crb1100283) | Discovery Peptides

In this study we have designed and synthesized the peptide-linked molecular beacons targeting U1, U2 snRNAs and U3 snoRNAs, as well as molecular beacons with a ‘random’ probe sequence. The specific oligonucleotide sequences of these MBs and the sequence of the NLS peptide are shown in , with the stem domain of a molecular beacon underlined. The design of these probes to target specific segments of small RNAs in the nuclei of living cells was based on previous fluorescence in-situ hybridization (FISH) studies in fixed cells with linear oligonucleotide probes, which were shown to be able to access the target (, , –). The ‘random’ beacon was designed as a negative control, with a 17-base probe sequence that does not have any exact match in the mammalian genome (, ). The probes used in this study are with deoxyribonucleotide backbone; the dye and quencher pairs (Cy3-BHQ-II and Cy5-BHQ-III) were selected to ensure effective quenching of the probes in their unhybridized state (stem-loop hairpin).