...MEDLINE-indexed journal Nuclear Receptor Signaling. Dr. McKenna edits the website of the Nuclear Receptor Signaling... elicit their actions on other tissues. At the Nuclear Receptor Signaling Atlas (NURSA), one of our primary...regulated at the cellular, tissue and organ level by nuclear receptors (NRs), their coregulators, and their ...their physiological endocrine ligands. Nuclear receptors and their ligands NRs represent the largest family...Consortium represents a unique funding model in nuclear receptor research, and has generated a set of comprehensive...address many important and fundamental questions in nuclear receptor and coregulator research. Our Q-PCR datasets...for example, document the expression patterns of nuclear receptors across multiple different tissues in ...

... 4D nucleome, the organization of the nucleus in space and time, and to understanding how nuclear organization...dCas9 gets a colorful makeover Even without its nuclease activity, catalytically dead Cas9 (dCas9) has ...loci, a major benchmark in characterizing the 4D nucleome. Future modifications and applications Ma et al...technique. The NIH Common Fund has made mapping the 4D nucleome a specific priority, as researchers work to increase... nucleome...

...updated April 16, 2020. Sensitive and specific nucleic acid detection is crucial for clinical diagnostics...biotechnological advancements. Many methods of nucleic acid detection however, either lack the sensitivity...sensitivity or the specificity to detect nucleic acids at low concentrations and/or are too expensive, time-consuming..., and inexpensive platforms to reliably detect nucleic acids at the atomolar level. The Zhang lab has...degradation to develop the method termed DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR) (Chen...signal that indicates the presence of your targeted nucleic acid. To increase the sensitivity of the assay,... In addition, Csm6, a CRISPR type-III effector nuclease, can be used in conjunction with Cas13 to amplify...

...reagents in a technique known as nucleofection, which forms pores in the nuclear membrane, allowing for entry...translation delays editing What are Cas9-gRNA ribonucleoproteins? One alternative approach, which avoids many...these complications, is to directly deliver a ribonucleoprotein (RNP), consisting of the Cas9 protein in complex...use RNPs, now what? Preparing the Cas9-gRNA ribonucleoprotein The first step in this type of CRISPR experiment...Schier laboratories. Delivering Cas9-gRNA ribonucleoproteins  An advantage of performing CRISPR experiments...fusion to a fluorescent protein is used to label nucleic acid targets.  While Cas9-gRNA RNPs show promise... efficient mouse genome editing by CRISPR ribonucleoprotein electroporation of zygotes." Journal of Biological...

...Considerations when choosing your Cas nuclease When choosing the Cas9 nuclease for your experiments, there are...but since then, scientists have adapted CRISPR nucleases for many more specific research needs. In this...determined by the PAM sequence recognized by the nuclease. For the most frequently used Cas9, spCas9 from... Figure 1: The PAM sequence that the Cas nuclease recognizes determines site accessibility. Cas9s... measure of the on-target activity of the Cas9 nuclease. This can be determined by measuring the indel...measure of on-target activity (right). Cas9 nucleases that exhibit enhanced site accessibility In order...engineered Cas9, SpCas9-NG, which required only the di-nucleotide PAM NG (Nishimasu et al., 2018). The David Liu...

... alpha PPARD Plasmids PPARD, FAAR, NR1C2, NUC1, NUCI, NUCII, PPARB peroxisome proliferator-activated receptor...Browse the nuclear receptor plasmid collection assembled in collaboration with the Nuclear Receptor Signaling... Plasmid Collections Nuclear Receptor Plasmid Collection Nuclear Receptor Plasmid Collection You...Wikipedia, accessed November 6, 2013. Author: Boghog2 Nuclear receptors are a class of transcription factors ...modulated by ligands. Two of the important regions of nuclear receptors are: 1) a ligand binding domain, which...activation or repression of target gene expression. Nuclear receptors play a key role in development and homeostasis...from around the globe, Addgene has assembled a Nuclear Receptor plasmid collection for the community. ...

...time using Custom Synthetic cr:tracrRNA/Cas9 Ribonucleoprotein (csRNP) delivery Epitope tagging of mammalian...with recombinant Cas9 protein to form the ribonucleoprotein complex (csRNP, Figure 1). We use in-house...cells recover for 2-5 days.Note: We use the Nucleofection method for delivery of csRNP reagents into cells...tagging in mammalian stem cells using Cas9 ribonucleoprotein. Elife 7. PubMed PMID: 29638216. PubMed Central... by improved design and delivery of gRNA, Cas9 nuclease, and donor DNA. J Biotechnol 241: 136-146. PubMed...

... participate when the organizers of the Canton Nucleic Acids Forum (CNAF) 2015 conference invited me to...Chinese scientists eager to hear current findings in nucleic acid science. I was delighted to see that the majority...plasmid requests from Addgene. Addgene - Canton Nucleic Acids Forum 2015 from Joanne Kamens, PhD Thoughts...structure.” That certainly summed up this conference—nucleic acid science still has many surprises for the researchers... nucleic acids...

...two major lobes, called NUC (for nuclease) and REC (for recognition). The NUC lobe contains the DNA scissors... of Cas9’s two major lobes, REC and NUC, and their subdomains. NUC includes the HNH and RuvC catalytic...conformation cleaves both DNA strands via the NUC lobe, as in Figure 6. NUC lobe, green; REC lobe, light blue; guide... domain (CTD) of the protein also belongs to the NUC lobe. Because its primary role is recognizing the...to, and their roles. Lobe Domain Role NUC HNH Cut DNA target strand RuvC Cut DNA ...’s most often shown in an “open” state, with the NUC and REC lobes laying open like a book (Figure 2)....conformation, as in Figure 3. Upon DNA binding, the NUC lobe recognizes the PAM site in the non-target DNA...

...Zinc Finger Consortium Nuclease Expression Vectors Zinc Finger Consortium: Nuclease Expression Vectors This...page lists plasmid vectors designed to express nucleases in different cell types. Zinc finger arrays can...page lists plasmid vectors designed to express nucleases in different cell types. Zinc finger arrays created...cloning sites. Vectors encoding wild-type FokI nuclease domains: pST1374 (for expression in mammalian ...in plants) Vectors encoding heterodimeric FokI nuclease domains: pMLM290 (for expression of a “+” heterodimeric... “-” heterodimeric FokI domain ( Sollu et al., Nucleic Acids Res ) in mammalian cells or zebrafish). pCLR2068...by Moore et al. PLoS One 2012 and Cade et al., Nucleic Acids Res 2012 ) pCLR2070 (for expression of a ...

...target genomic DNA sequence and a nuclease (e.g.—the Cas9 endonuclease) that cleaves the target DNA. The...CRISPR-Cas9 nucleases with altered PAM specificities Dimeric CRISPR RNA-guided FokI Nucleases (RFNs) CRISPR-Cas...Joung Lab CRISPR-Cas/RNA-Guided Nuclease (RGN) Plasmids Genome...Plasmids Joung Lab CRISPR-Cas/RNA-Guided Nuclease (RGN) Plasmids You may also like... CRISPR Guide CRISPR...gRNA Design Tools CRISPR Blog Posts RNA-Guided Nucleases ( RGNs ), based on naturally occurring Type II...II CRISPR-Cas systems, are programmable endonucleases that can be used to perform targeted genome editing...target sites and to facilitate design of short oligonucleotides needed to construct gRNA expression vectors...

...pAAV-mDlx-NLS-mRuby2 112677 AAV1, AAV2 pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP 27056 AAVrg pAAV-Ef1a-DIO EYFP...pAAV-Ef1a-fDIO mCherry 112677 AAV2 pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP 50457 AAV2 pAAV-hSyn-DIO-EGFP...

...forebrain GABAergic interneurons. pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP (112677-AAV5 and 112677-AAVrg)...H134R)-mCherry (26976-AAV5) pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP (112677-AAV5) Calcium Sensors...switch that expresses nuclear mCherry in the absence of Cre, and expresses nuclear EGFP in the presence...

...Karl Deisseroth 112677 AAV2 pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP Brandon Harvey 50457 AAV2 pAAV-hSyn-DIO-EGFP...

...pCaggs-NLS-PAmCherry1-GSS-EGFP Nucleus NLS mGold Francois St-Pierre 14893 pEBFP2-Nuc Nucleus NLS EBFP2 Robert Campbell...pmKalama1-Nuc Nucleus NLS mKalama1 Robert Campbell 73205 pcDNA3.1-NLS-myc-dL5-2xG4S-mCer3 Nucleus NLS (from...Cytosol, nucleoplasm None dL5 FAP; mCerulean3 Marcel Bruchez 36206 pmTurquoise2-NES Non-nucleus Nuclear Export...Non-nucleus Nuclear Export Sequence mScarlet-I Dorus Gadella 85061 pmScarlet-H_NES_C1 Non-nucleus Nuclear...3xnls-mNeonGreen Nucleus 3xNLS mNeonGreen Dorus Gadella 98817 3xnls-mTurquoise2 Nucleus 3xNLS mTurquoise2...pEGFP-C1-Fibrillarin Nucleolus Fibrillarin EGFP Sui Huang 36207 pmTurquoise2-H2A Nucleus H2A mTurquoise2 Dorus...pmScarlet-i_H2A_C1 Nucleus H2A mScarlet-I Dorus Gadella 85052 pmScarlet-H_H2A_C1 Nucleus H2A mScarlet-H Dorus...

...PHP.eB, PHP.S Boyden 112677 pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP EF1a NLS-mCherry or nls-EGFP Cre...

...Fluorescent Protein Guide NLS Nuclear Localization pENTR4-myc-nuc - pEntry vector that...that adds a C-terminal nuclear localization signal FNpcDNA3 - N-terminal ... FLAG and SV40 nuclear localization signal Myr Membrane Localization pWZL-Neo-Myr-Flag-DEST - ...

...EYFP Control Gradinaru 112677 pOTTC1032 - pAAV EF1a Nuc-flox(mCherry)-EGFP EF1a Color-flipping switch. Expresses...GCaMP6f-P2A-nls-dTomato Syn GCaMP6f and physically separate nuclear dTomato Calcium sensor Ting 51083 AAV-EF1a-DIO-...AAV-EF1a-DIO-GCaMP6f-P2A-nls-dTomato EF1a GCaMP6f and physically separate nuclear dTomato, Cre-dependent Calcium sensor Ting 83899...GCaMP6s-P2A-nls-dTomato Syn GCaMP6s and physically separate nuclear dTomato Calcium sensor Ting 100853 pAAV.Syn.Flex.NES-jRGECO1a.WPRE.SV40...

...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...