We narrowed to 417 results for: NUC

...a zinc finger (ZF) nuclease. Zinc fingers recognize triplets and the FokI nuclease operates as a dimer...sites. (B) A TAL effector nuclease (TALEN). Similar in principle to the ZF nuclease, the components of the...the array recognize individual nucleotides. For over ten years, zinc finger (ZF) arrays have been the go-to...number of zinc fingers that recognize various nucleotide triplets. With some trial and error, ZF arrays...years of research has shown that when fused to a nuclease or activation domain, a ZF array is an effective...without their limitations, however. Not every nucleotide triplet has a corresponding zinc finger, and ...Bogdanove at Iowa State University published the nucleotide recognition code of the TAL effectors, which ...

...10aa-H2B-WPRE3-BGHpA AAV vector for nuclear SYFP2 labeling in neurons. Useful for nuclear isolation and scRNA-seq ...pAAV-hDlx-GiDREADD-dTomato-Fishell-5 Gi-DREADD (P2A) nuclear dTomato expression in forebrain GABA-ergic interneurons...pAAV-hDlx-GqDREADD-dTomato-Fishell-4 Gq-DREADD (P2A) nuclear dTomato expression in forebrain GABA-ergic interneurons...

...genome engineering technologies like zinc finger nucleases (ZFNs) and TALENs. CRISPR: An Adaptive Immune ...Through sequence homology, these crRNAs guide a Cas nuclease to the specified exogenous genetic material, which...the individual crRNAs. (3) The tracrRNA and Cas9 nuclease form a complex with each individual, unique crRNA... alternative PAM sites . Cpf1 , another CRISPR nuclease, recognizes a 5’-TTN PAM, improving targeting ...Cascio VM, Joung JK. 2014. Improving CRISPR-Cas nuclease specificity using truncated guide RNAs. Nat Biotechnol...Shinagawa H, Makino K, Amemura M, Nakata A. 1987. Nucleotide sequence of the iap gene, responsible for alkaline... E. 2012. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science . 337...

... CRISPR-mediated nucleic acid cleavage. We have harnessed the type II CRISPR nuclease system to facilitate...nterspaced S hort P alindromic R epeats) is a microbial nuclease system involved in defense against invading phages...by co-expressing the S. pyogenes Cas9 (SpCas9) nuclease along with the guide RNA. For most applications... have found that adding an additional guanine nucleotide to the 5’ of 20-bp guide can increase targeting... genes. It consists of three components: A nucleolytically inactive Cas9-VP64 fusion A sgRNA incorporating... have found that adding an additional guanine nucleotide to the 5’ of 21-bp guide can increase targeting...GFP-KASH fusion facilitates FACS sorting of cells and nuclei. Return to top AAV - Cas9 mouse CRISPR-Cas9 is ...

...NF1 Neurofibromin 1 NFE2L2 Nuclear factor, erythroid 2 like 2 NFKB1 Nuclear factor of kappa light polypeptide...Ral guanine nucleotide dissociation stimulator RAPGEF RAPGEF1 RAPGEF2 Rap guanine nucleotide exchange factor...GTPase and is a member of the G protein (guanine nucleotide binding protein) family. GTPases serve as molecular...activating protein 35 ARHGEF2 Rho/Rac guanine nucleotide exchange factor 2 CCND CCND1 CCND2 CCND3 Cyclin... RASGRF1 RASGRF2 Ras protein specific guanine nucleotide releasing factor RASGRP RASGRP1 RASGRP2 RASGRP3...scaffold protein SOS SOS1 SOS2 SOS Ras/Rho guanine nucleotide exchange factor SPRED SPRED1 SPRED2 SPRED3 Sprouty...TSC1 TSC2 Tuberous sclerosis VAV1 Vav guanine nucleotide exchange factor 1 YAP1 Yes associated protein...

... the SARS-CoV-2 nucleocapsid protein. Brian Geiss 211757 Anti-SARS-CoV-2 Nucleocapsid Protein [mBG86] ...Antibody Description PI 211756 Anti-SARS-CoV-2 Nucleocapsid Protein [mBG17] Clone mBG17, recombinant mouse...monoclonal antibody targeting the SARS-CoV-2 nucleocapsid protein. Brian Geiss Plasmids SARS-CoV-2 Plasmids...spike protein, S; small envelope protein, E; nucleocapsid protein, N; and membrane protein, M), including...Cas12a and Cas13a plasmids, to reliably detect nucleic acids SARS-CoV-2 main protease (Mpro) activity ...Plasmid Description Industry PI Addgene blog Finding nucleic acids with SHERLOCK and DETECTR No Llamas Required...

...40822 EGFP-alphasynuclein-WT SNCA GFP CMV Parkinson's David Rubinsztein 40823 EGFP-alphasynuclein-A53T SNCA... pHM6-alphasynuclein-WT SNCA His, HA CMV Parkinson's David Rubinsztein 40825 pHM6-alphasynuclein-A53T ... Alpha-synuclein SNCA T7 Parkinson's Michael J Fox Foundation MJFF 51486 pET21a-alpha-synuclein SNCA T7...Alpha-synuclein SNCA T7 Parkinson's Michael J Fox Foundation MJFF 89074 pD454-SR rat Alpha-synuclein SNCA...Alpha-synuclein SNCA T7 Parkinson's Michael J Fox Foundation MJFF 89076 pD434-SR rat Alpha-synuclein SNCA...89470 VN-alpha Synuclein SNCA Venus CMV Parkinson's Bradley Hyman 89471 alpha Synuclein-VC SNCA Venus ...89483 VN-beta Synuclein SNCB Venus CMV Lewy body dementia Tiago Outeiro 89484 beta Synuclein-VC SNCB Venus...

...mammalian cells. Like Cas9 nucleases, Cpf1 family members contain a RuvC-like endonuclease domain, but they lack...contains a RuvC-like endonuclease domain, but it lacks Cas9’s other HNH endonuclease domain, indicating ...functional CRISPR nuclease that could be adapted for genome editing. The use of a different nuclease could potentially...lack Cas9’s second HNH endonuclease domain. Cpf1 cleaves DNA in a staggered pattern and requires only ... Figure 1: Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Figure from ... Cas9-based system, require a single-component nuclease to mediate cleavage rather than the multi-subunit...employed by class 1 systems. A putative new class 2 nuclease, Cpf1 (CRISPR from Prevotella and Francisella)...

...mutated to inactivate the protein’s nucleolytic capability, and this nuclease-null or “dead” Cas9 (dCas9) variant...relies on two distinct Cas9 variants—nuclease-competent or nuclease-null—in order to effect different perturbations...imagine transfecting cells with both the nuclease-positive and nuclease-null variants of Cas9 along with the...(gRNA). The gRNA conventionally contains a 20-nucleotide sequence that is complementary to the target ...Alex Chavez Figure 1: Schematic of native (nuclease-competent) Cas9 interacting with either a full-length... many cases this is due both to lower relative nuclease efficiencies as well as to more stringent targeting... An alternative approach is to modulate Cas9’s nuclease activity by modifying the gRNA with which it is...

...serve as slaves.  Meganucleases In 1989 the first ever DNA-editing enzymes, meganucleases, were discovered...created by genetics.    Zinc finger nucleases Zinc finger nucleases (ZFNs) are artificial restriction enzymes...instead of recognizing nucleotide triplets, the components recognize individual nucleotides. This imparts TALENs...The major drawback of all the endonuclease-based gene editors (meganucleases, ZFNs, TALENs, and CRISPR-Cas...altering the cleavage specificity of a homing endonuclease. Nucleic Acids Research 30:3870–3879 . https://doi.org...in 1990, just a year after the discovery of meganucleases. The storyline involves the resurrection of ... to a DNA-cleavage domain (FokI). Zinc finger nucleases were first designed in 2005. The DNA-binding domain...

..."The polynucleotide ligase and RNA capping enzyme superfamily of covalent nucleotidyltransferases." Current...mispaired bases, and one-nucleotide gaps can be sealed by T4 DNA ligase." Nucleic acids research 15.21 (...al. "Ligase-based detection of mononucleotide repeat sequences." Nucleic acids research 27.24 (1999): e40...accelerate characterization of nucleic acid metabolic enzymes." Nucleic acids research 44.2 (2015): e15...DNA ligases. These applications detect specific nucleotide sequences with high specificity and quantitative...quantitative accuracy and even include profiling single nucleotide polymorphisms (SNPs) (5,12,13). For example, ...David H. Gelfand. "Cloning, overexpression and nucleotide sequence of a thermostable DNA ligase-encoding...

...that occurs during nuclear envelope breakdown (without the nucleus, there’s no nuclear localization to monitor...further fused to a bipartite nuclear localization signal (bNLS) and a nuclear export signal (NES), both ...the nucleus depending on its phosphorylation status. The relative ratio of cytoplasmic vs. nuclear fluorescence... because KTRs shuttle between the nucleus and the cytoplasm, nuclear export and import rates can influence...Prior to phosphorylation, the KTR localizes to the nucleus, but following phosphorylation bNLS activity is...kinase activity that occurs in the cytoplasm and/or nucleus, while FRET-based reporters can be used to measure... rates have negligible effect on KTRs, however nuclear translocation rates are another variable to consider...

...Transformed cell lines Cargo Nucleic acids or protein Nucleic acids Nucleic acids Throughput High High... Dividing or nondividing Cargo Nucleic acids Nucleic acids Nucleic acids Throughput Low Low Low ...Electroporation / Nucleofection Microinjection Cells Many types Single cells Cargo Nucleic acids or protein...deliver a donor plasmid or single-stranded DNA oligonucleotide, so you will need to make sure your expression...reagents can be delivered via transfection, nucleofection, viral transduction or injection as either protein...generating transgenic embryos. Cas9-gRNA ribonucleoprotein complexes Purified Cas9 protein and in vitro...protein Nucleic acids or protein Throughput High Very low Efficiency High Very high Difficulty Very...

...have two endonuclease domains: the n-terminal RuvC-like nuclease domain and the HNH-like nuclease domain...Cas9 genome editing systems use a non-specific endonuclease (Cas9 or closely related Cpf1) to cut the genome...genome and a small RNA (gRNA) to guide this nuclease to a user-defined cut site. After reading this post... have increased targeting specificity). Cas9 endonuclease functions While native CRISPR/Cas systems have...foreign DNA as well as the RNA guides required for endonuclease function, when used for genome engineering, ...the only CRISPR protein required is the Cas9 endonuclease or a variant thereof. This individual protein...) of a protospacer adjacent motif (PAM) Cas9 endonuclease binding to the target genomic locus is mediated...

...vulnerable to attack by both cytoplasmic and nuclear nucleases, as they will be recognized as foreign RNA...to simply swap out several of the ribonucleotides for deoxynucleotides. A partially DNA gRNA is surprisingly...efficiency. In the same vein, locked nucleic acids or bridged nucleic acids can also be introduced in the...hydroxyl group of the ribose to protect it from nucleolytic attack. Another stabilizing sugar modification...modifications to consider include amide linkages, unlocked nucleic acids, and a constrained ethyl.  Can’t decide which...which improves mismatch discrimination and is more nuclease-resistant, resulting in less off-target events...events. While both have similar advantages, bridged nucleic acids with N-methyl substitutions are slightly ...

...TAL effector nuclease (TALEN) facilitates efficient knockout of HIV co-receptor CCR5. Nucleic Acids Research...researchers instead turn to so-called “mismatch nuclease” assays (e.g. Surveyor® or T7E1) for mutation ...Digital PCR has several advantages over mismatch nuclease assays that will be elaborated below. Digital...advantages over the commonly employed mismatch nuclease assays. Sensitivity: Less than 20 ng of total...mutant targets have been reported for mismatch nuclease assays (2, 6, 7). Ability to distinguish between...functional knockout of your target gene. Mismatch nuclease assays are completely “blind” to this very important...gene-editing frequencies (GEF-dPCR) mediated by designer nucleases. Nat Protoc 11, 598–615 (2016). PubMed PMID: 26914317...

...10-6 mutations per nucleotide per generation vs. 10 x 10-10 mutations per nucleotide per generation of ...provide a means to continuously diversify all nucleotides within a user-defined genomic region. EvolvR,...users to achieve targeted diversification of all nucleotides within a defined region of a genome. Require...turbidostat. Only mutate particular types of nucleotides. Both the Bassik Lab and the Chang Lab have created...Diversification is limited due to the integration of oligonucleotide libraries of a discrete size at the target ...extends it from the 3’ end, while its native endonuclease activity degrades the displaced strand.   ...right), occasionally introducing mutations (red nucleotide). Created with BioRender.com. The initial...

...the order of nucleotides in a string of DNA (7). Sanger sequencing uses modified nucleotides, called dideoxynucleotides...synthesis reaction along with all four regular nucleotides results in the synthesis of DNA strands of all...contain radioactive bands that correspond to every nucleotide in your DNA sequence, based on which ddNTP was...Compiling the identity and location of a given nucleotide, a researcher (or a simple computer program) ...Building upon this technology, radiolabelled nucleotides were replaced with fluorescently labeled ones...fluorescence activity as fluorescently labeled nucleotides are incorporated, a method called pyrosequencing...nanopore-based DNA sequencing which reads the nucleotides of a DNA strand as it slips through a tiny pore...

...genome, the mouse genome is made up of 3 x 109 nucleotides (nt), and encodes 23,000 or so genes. It would...about 125 nt that specifies the target, the Cas9 endonuclease that creates the DNA double-strand break (DSB...DSB) at the target site, and a donor oligonucleotide or plasmid as the repair material if needed (for ...create a mouse model, the gRNA, Cas9, and donor oligonucleotide or plasmid components are brought together ...together and microinjected into either the pronucleus or the cytoplasm of fertilized mouse eggs. Or, to avoid... technique called Genome editing via Oviductal Nucleic Acids Delivery, or GONAD (Gurumurty et al., 2016.... Alternatively, the gRNA, Cas9, and donor oligonucleotide can be electroporated into the mouse zygote...

...menu. Typically, we use the default nucleotide database “Nucleotide collection (nr/nt)” as it contains ...programs available, we primarily use Standard Nucleotide BLAST (blastn), Standard Protein BLAST (blastp...ORFs) and determine the potential effect of any nucleotide discrepancies. The blastp and blastx programs...below. Optimizing blastn searches On the Standard Nucleotide BLAST page, the first decision to make is whether...BLAST sequence database. If you know the expected nucleotide sequence, check the “Align two or more sequences...Subject Sequence box that appears. Aligning two nucleotide sequences is probably the fastest BLAST search...noted any potential mismatches or gaps in the nucleotide sequence, you can run a Translated BLAST (blastx...