We narrowed to 417 results for: NUC

...with and without targeted nucleases). AAV genome editing without a nuclease Although the mechanism of...cells. AAV genome editing combined with targeted nucleases In an effort to combine the respective advantages...introduction of a double strand break by using a targeted nuclease. Asuri et al, for example, used zinc fingers to...devised methods to deliver CRISPR components as ribonucleoproteins (RNPs) that can be supplied separately from...knockouts using recombinant adeno‐associated viruses." Nucleic acids research 32.1 (2004): e3-e3. PubMed PMID:...endogenous genes in diverse human cell lines." Nucleic acids research 36.19 (2008): e127-e127. PubMed ... and David V. Schaffer. "Combining engineered nucleases with adeno-associated viral vectors for therapeutic...

...3;108(18):7351-6. Amy Palmer Zinc Cytosolic or Nuclear zinc-binding FRET biosensor (Zap) New alternately...bacterial or mammalian expression, targeted to nucleus, cytoplasm, or mitochondria A genetically encoded...Photosynthesis-dependent H 2 O 2 transfer from chloroplasts to nuclei provides a high-light signalling mechanism. Nat...cellular concentrations of nicotinamide adenine dinucleotides. eLife. 2018 May 29;7. pii: 32638. Kai Johnsson...GRAB-HTR6 A serotonergic axon-cilium synapse drives nuclear signaling to alter chromatin accessibility. Cell... protein pH biosensor to detect concentrative nucleoside transport. J Biol Chem. 2009 Jul 31;284(31):20499...

...are only able to enter the nucleus during mitotic breakdown of the nuclear envelope. Lentiviruses can ...components are small enough to enter the nucleus through the nuclear pore complex. This feature may be useful...retroviral particle containing Matrix, Capsid, and Nucleocapsid components. Pol Precursor protein containing... Psi (Ψ) RNA target site for packaging by Nucleocapsid. WPRE Woodchuck hepatitis virus post‐transcriptional...stimulates the expression of transgenes via increased nuclear export. LTR Subcomponents: -U3 -R -U5 LTR...

...implanted into a female animal for gestation. Nucleases that induce double-stranded DNA breaks, such as...inserted gene may have errors introduced to the nucleotide sequence during the DNA repair process. The addition...cells, which are then injected into a blastocyst. Nucleases can also be used to engineer knockout mice. ZFN...non-homologous repair at the site of DNA damage. These nucleases are microinjected into a zygote, which is then...introduce point mutations. While using targeted nucleases to generate an engineered mouse model has advantages...targeting vector alone, some challenges still exist. Nucleases may cause off-target effects on regions of the... random process, zygotes edited using the same nuclease may not result in genetically identical mice. ...

...fragment is actively shuttled out of the nucleus due to the nuclear export sequence. Treatment with rapamycin...DNA recognition domain and a mostly c-terminal nuclease domain. Feng Zhang's group utilized the bi-lobed...FKBP-2xNLS dimerization and net influx into the nucleus, where the functional Cas9 molecule can cleave ... systems such as TAL effectors or Zinc Finger Nucleases, the Gersbach lab has modified the RNA-guided ...by the use of sequence-specific enzymes (e.g. nucleases, recombinases) which irreversibly excise or invert...bound by both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP...rebind because they are transported back to the nucleus (thanks to a NLS on these proteins). The RFP proteins...

... can be assembled. For example, because CpG dinucleotides are underrepresented in most vertebrate genomes...and MluI recognition sites, which contain CpG dinucleotides in their 6-mer sequences, are rarely found in...for cloning genes from organisms with high CpG dinucleotide representation. The specific restriction enzymes...Kaiser AD, Kornberg A. Enzymatic synthesis of deoxyribonucleic acid. VIII. Frequencies of nearest neighbor...neighbor base sequences in deoxyribonucleic acid. The Journal of biological chemistry. 1961. March;236:864–...Trautner TA, Kornberg A. Enzymatic synthesis of deoxyribonucleic acid. XI. Further studies on nearest neighbor...neighbor base sequences in deoxyribonucleic acids. The Journal of biological chemistry. 1962. June;237:1961...

...is?  To assess the concentration and purity of nucleic acids, such as plasmids, the go-to method is to...three key wavelengths: 230 nm, 260 nm, and 280 nm. Nucleic acids exhibit a characteristic absorption spectra...Beer–Lambert law, the instrument can calculate a sample’s nucleic acid concentration based on the sample’s absorption... reagent contaminants The reagents used during nucleic acid purification can leave behind a variety of...G and Cantor E. A Practical Guide to Analyzing Nucleic Acid Concentration and Purity with Microvolume ...Loughrey S and Matlock B. Detection of Protein in Nucleic Acid Samples Using the NanoDrop One Spectrophotometer...Loughrey S and Matlock B. Detection of phenol in nucleic acid samples using the NanoDrop One spectrophotometer...

.... Among the most well-known CRISPR-associated nucleases are Cas9 and Cas12a, the heavyweights in the genome... has become a key focus in recent years. TnpB nucleases represent an exciting class of compact genome ...proteins. Functionally, while Cas9 contains two nuclease domains (HNH and RuvC-like), and Cas12a has one... the transposon's right-end element, and a 20-nucleotide (nt) guide sequence that is complementary to ...formation recruits TnpB to the site and activates its nuclease function. Once bound, TnpB cleaves the DNA at ...accommodate the large coding sequence of commonly used nucleases like Cas9. This is where TnpB offers a clear advantage...increasing the overall size), using TnpB as the core nuclease could help keep the total construct size smaller...

...precision with dual-nuclease Cas9-Cas9 chimeras Scot Wolfe’s lab has developed dual-nuclease Cas9-Cas9 chimeras...CRISPR tools for your research.     Improved nucleic acid sequence detection Feng Zhang’s lab recently...editing in plants While there have been many Cas nucleases developed for plants, it has been difficult to...to compare the performance of different nucleases. Nicola Patron’s lab compared the efficiency and specificity...multiple wild-type and engineered variants of Cas nucleases. In doing so, they also created a modular toolkit...

...intracellular RNA dynamics. Aptamers are 10-100 base nucleic acid oligos that bind with high affinity to small...describes the enrichment process used to evaluate oligonucleotides for binding affinity to a specific target,...of Spinach. In doing so, they identified the nucleotides that contribute to the instability of Spinach...also smaller than any form of Spinach - only 49 nucleotides in length - and is better able to maintain its...that developed Corn. Aptamers using unnatural nucleotides are also being developed which could help aptamers... Panchapakesan, Shanker Shyam S., et al. "Ribonucleoprotein purification and characterization using RNA...superfolding Spinach2 reveals the dynamic nature of trinucleotide repeat–containing RNA." Nature methods 10.12...

...methylation-dependent endonuclease (i.e. DpnI), and bacteria are transformed with the nuclease-resistant nicked...), 3’->5’ exonuclease activity (increases amplification fidelity), and no 5’->3’ exonuclease activity ...technique that can be used to introduce specific nucleotide substitutions (or deletions) in a tailored manner... cannot be used for site directed mutagenesis. Nuclease To remove the template DNA (unmodified plasmid...DNA polymerase for site-directed mutagenesis." Nucleic acids research (2014): gku1189. PubMed PMID: 25399421...

...Exon-skipping antisense oligonucleotides to correct missplicing in neurogenetic diseases. Nucleic Acid Ther. 2014...for DMD treatment. Clinical trials have used oligonucleotide exon skipping (OEN) to remove mutated exons... dystrophin transcript. Unfortunately, the oligonucleotides only modestly improve muscle function, and...Dystrophin and genome editing Since complex oligonucleotide treatment comes with many challenges, researchers...depositor Charles Gersbach used paired zinc finger nucleases to remove exon 51 in DMD patient myoblasts. They...through genomic excision of exon 51 by zinc finger nucleases. Mol Ther. 2015a Mar;23(3):523-32. PubMed PMID...

...EF1alpha addition of an mCherry reporter variant nuclear reporters via an H2B fusion shuttle plasmid for...design tools and ordered as standard desalted oligonucleotides with the appropriate overhangs: The plasmids...quick ligation of annealed and phosphorylated oligonucleotides containing the guide target sequence: Up to...

...since potential target sites occur ~1 in every 8 nucleotides, restricting gRNAs to 5 – 65% of the protein ...Anzalone et al., 2020). is not limited to single nucleotide transitions but still requires a nearby PAM, ...activation) and CRISPRi (inhibition) technologies – a nuclease-dead Cas9 (dCas9) is directed near the promoter...the generation of truly isogenic cell lines than nuclease-mediated off-target effects.” Further, large-scale... Liu DR (2020) Genome editing with CRISPR–Cas nucleases, base editors, transposases and prime editors....CRISPR/dCas9-mediated transcriptional repression. Nucleic Acids Res 44:e141–e141 . https://doi.org/10.1093...Optimization of scarless human stem cell genome editing. Nucleic Acids Research 41:9049–9061 . https://doi.org/10.1093...

...previous strategy for multiplexing gRNAs used the RNA nuclease Csy4 from Pseudomonas aeruginosa to cleave 4 gRNAs...with Cas9 and the polycistronic transcript. As nucleases may have off-target effects and/or toxicity, it...cleaved precisely as predicted with no addition of nucleotides to the 5’ spacers. The 3’ ends were modified ...tRNA-gRNA-tRNA constructs. gRNAs with any 5’ leading nucleotide could be processed, a distinct advantage over...over other methods that favor certain nucleotides at the 5’ end. PTG construct schematics. Scissors indicate...at Addgene Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing. Tsai SQ, Wyvekens...

...recognizable sub-cellular localization (membrane, nucleus, etc.) are also available Non-toxic to living cells...pSNAPf-H2B Control Plasmid SNAP-tag Control for nuclear localization 101123 pSNAPf-ADRβ2 Control Plasmid...pCLIPf-H2B Control Plasmid CLIP-tag Control for nuclear localization 101130 pCLIPf-Cox8A Control Plasmid...

..., additional nuclear localization signals, and two mutations in Cas9 to enhance nuclease activity; can...or Cas9 domains PE7 — addition of RNA-binding exonuclease protection factor La to PEmax to enhance pegRNA...

...binding proteins, this method can characterize nucleic acid-nucleic acid interactions at a genomic locus (Fujita...numerous discoveries have been made using the Cas9 nuclease in the absence of genomic alterations. These studies...and allowed the Xu lab to identify protein and nucleic acid binding partners at numerous genomic loci ...enCh IP technology using recombinant CRISPR ribonucleoproteins." Genes to Cells 21.4 (2016): 370-377. PubMed..., et al. "A programmable dual-RNA–guided DNA endonuclease in adaptive bacterial immunity." Science337.6096...

...Protospacer Adjacent Motif (PAM sequence). The 3-5 nucleotide PAM sequence serves as a binding signal for Cas9...genome engineering was Cas12a (formerly Cpf1), a nuclease that generates double strand breaks in target ...Genome-wide analysis reveals specificities of Cpf1 endonucleases in human cells. Nat Biotechnol 34:863–868 . ...Aryee MJ, Joung JK (2015) Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature 523:481... Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells. Nat Biotechnol 34:869–874 . https..., Zhang F (2015) Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell 163:759...

...Note: hypoxanthine (shown) is the nucleobase component of the nucleoside inosine. Created with BioRender.com...newly-deaminated inosines. Pro tip! The nucleoside inosine consists of the nucleobase hypoxanthine + a ribose sugar...initial editor AXBE, for A → X base editor (X = any nucleotide). To create a more specific A → C base editor...invented.   Abasic site A DNA site with the nucleobase removed. Also called an apurinic / apyrimidinic...