We narrowed to 735 results for: RAN-1

..., or the beginning of the coding sequence. Figure 1: Overview of CRISPR interference. Browse, sort, or...CRISPR plasmids for transcriptional repression of target genes. CRISPR...Catalytically dead Cas9 (dCas9), or dCas9 fused to a transcriptional repressor peptide like KRAB, can knock down...down gene expression by interfering with transcription. This interference is most efficient when the gRNA...

...for any application TMN200 (200 mM sodium chloride, 1 mM magnesium chloride, 20 mM Tris at pH 8.0) Suitable...adeno-associated viral vector. Hum Gene Ther Methods . Feb;25(1):72-82. PMID: 24191859 Ye, et al. 2015. Safety and...enhanced transduction of the retina following subretinal administration and enhanced transduction and spread...cite Boye, et al. 2016. Impact of Heparan Sulfate Binding on Transduction of Retina by Recombinant Adeno-... from the University of Florida for enhanced transduction of the eye and other tissues. As part of our...Y444F) serotype is demonstrated to have enhanced transduction of the retina following intravitreal injection... AAV2(Y444F) serotype also displays enhanced transduction of the trabecular meshwork following intracameral...

...Expression mEGFP 488 507 34 6 Monomer (A206K) pmEGFP-1 - Mammalian Expression mEGFP-N1 - Mammalian Expression...TurboRFP-pBAD - Bacterial Expression tdTomato 554 581 95 4.7 1 h Tandem-dimer pCSCMV:tdTomato - Mammalian Expression...-pBAD - Bacterial Expression mRFP1 584 607 13 4.5 1 h Monomer pcDNA3-mRFP - Mammalian Expression pMXs-...Mammalian Gateway Destination Vector pENTR4-HaloTag (w876-1) - Gateway Entry Vector pCS2+/C-Halo , pCS2+/N-Halo...pSNAPf-C1 - Mammalian Expression pENTR4-SNAPf (w878-1) - Gateway Entry Vector pCS2+/C-SNAPf , pCS2+/N-SNAPf... Monomer pET mOrange LIC cloning vector (u-mOrange) - Bacterial Expression pCS2+8CmOrange - Zebrafish/...urchin pCS2+8NmOrange - Zebrafish/Xenopus/Worm/Sea urchin mOrange-N1 - Mammalian Expression mOrange-C1 - Mammalian...

...enzyme and guide RNA) for your experiment (Figure 1). You will decide how to express Cas9, the delivery...finally how to validate your genetic edit. Figure 1: Flow chart describing the general framework of a ...population. Some cells may remain wild type due to either (1) a lack of gRNA and/or Cas9 expression or (2) a lack...Biology , 1239 , 197–217. https://doi.org/10.1007/978-1-4939-1862-1_10 PMID: 25408407 Dixit, A., Parnas, O...fused to transcriptional activators that need to be within a given range of the transcription start site... interfere with transcription elongation. CRISPR Activation : Target the transcription start site. CRISPRa... perform by chemical transfection or electroporation. Expression can be transient, or you can generate...

...Buenrostro JD, Regev A, Zhang F. Cell 2023 Jan 5; 186(1):209-229. PubMed (Link opens in a new window) . TF...Browse the Zhang lab’s comprehensive human transcription factor collection, available at Addgene. The...The collection can be used for screening transcription factors for cellular programming or cancer drug ...Library The MORF library is a comprehensive human transcription factor collection that contains 1,836 genes ...collection, can be purchased at Addgene. Each transcription factor isoform has a unique 24 bp barcode that...that simplifies identification of the transcription factor isoform in pooled screens. This collection ...collection and library can be used for screening transcription factors for applications such as cellular programming...

...the dCas9-activator to your specific locus. Figure 1: Overview of CRISPR activation. Created with BioRender.com...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ...

...in mammalian cells or zebrafish). pMG10 (a '“2-in-1” ZFN cassette for expression of both a “+” and “-”...different cell types. Zinc finger arrays can be transferred in-frame to these vectors. Plasmid...either OPEN or modular assembly can be easily transferred in-frame to any of these vectors using unique...

...research and innovation programme and EFPIA. Figure 1: Representation of RESOLUTE mission. The combination...CRISPR knockout library targeting human transmembrane transporters. Feedback Please send your feedback to...carrier proteins (SLCs) are the largest family of transporters in the human genome with approximately 450 identified... Medicines Initiative 2 Joint Undertaking under grant agreement No 777372. This Joint Undertaking receives...allow a fast and efficient sub-cloning into a wide range of Gateway-compatible pDEST vectors. ID Plasmid ...overexpression library of human SLC proteins. Human Transporter Knockout Libraries 213695 221409 221410 Superti-Furga...

...Biology, 2(9), 537–541. DOI: 10.1021/sb400021j Figure 1: Multiplexing CRISPR knockdown to modulate expression...controlling the number of mismatches we can obtain a range of expression levels. This collection comprises ...Together, these plasmids allow to explore the whole range of expression levels for these two reporters simultaneously... S. (2018). Tuning dCas9’s ability to block transcription enables robust, noiseless knockdown of bacterial...

...gene activation CRISPR/Cas9 Plasmid Variants Figure 1: Schematic of the CRISPR Collection. Plasmids 159654... and 159655 are single guide expressing double stranded break constructs. Plasmids 171098 and 175570 allow...

...activation-induced cytidine deaminase (AID). Figure 1: Overview of CRISPR base editing. Browse, sort, or...generate single base pair edits without double stranded breaks. Cytosine base editors are created by fusing...creating a C to T change (or a G to A on the opposite strand). Likewise, adenine base editors have been engineered...

...) a T ranscriptional R epressor D omain (TRD) the TRD contains a Nuclear receptor Co-Repressor 1/Silencing...affecting females at an incidence of approximately 1:10,000. It is characterized by apparently normal development...reactivation, RNA editing, gene replacement therapy, RNA trans-splicing, and protein replacement to reverse the...cases Rett syndrome can be inherited. In females, random X inactivation results in approximately half of...disease severity. The MECP2 protein is a global transcriptional regulator of thousands of genes and studies...studies have suggested roles in transcriptional repression, activity dependent de-repression, chromatin remodeling...DNA and recruit known co-repressors or other transcriptional modifiers suggests the major function of MECP2...

...fluorescent proteins in budding yeast. Sci Rep, 9 (1):2234. https://doi.org/10.1038/s41598-019-38913-z ...von Wirén, N., Frommer, W. B. (2007). A cytosolic trans-activation domain essential for ammonium uptake....

...for brain circuit mapping and connectomics. Figure 1: Reconstruction of a sparse set of neurons across ...opens in a new window) , in collaboration with The Francis Crick Institute (Link opens in a new window) , ..., H.G.J., Leible, D., Leung, K.S., Magno, C.A., Meeran, A., Michalska, J.M., Rieger, F., Wang, C., Wu,...

...1774 for neomycin selection. Bob Weinberg 10676 pMKO.1 GFP MoMLV U6-driven plasmid for shRNA expression with...into multiple plasmids for safety: Transfer plasmid — contains transgene, sgRNA, or shRNA of interest flanked... the transgene fused to eGFP. Hans Clevers 18760 MSCV IRES Luciferase MSCV Plasmid for transgene expression...and transfer plasmids. Viral... Viral Plasmids Gamma-Retroviral Plasmids Gamma-Retroviral Plasmids Transfer Plasmids...encoded on RNA. The genome includes a reverse transcriptase and an integrase to insert the viral genome ...can take advantage of this to stably integrate transgenes of interest into a variety of cell types. Wild-type...

...and interrogate with conventional reagents. Figure 1: Axon guidance cues and receptors regulate the development...researchers with powerful tools to investigate neurotransmission, synapse formation, neural connectivity, plasticity...pairs bridge the presynaptic and postsynaptic membranes at neuronal synapses. Image credit: IPI Browse... top Integrins Integrins are a family of mechanotransduction receptors, comprised of alpha-beta subunit...

... Caltech and can help guide your decision. Figure 1: Decision tree for choosing your AAV capsid. Your ...tdTomato Control Boyden 83900 pAAV-mDlx-GFP-Fishell-1 Dlx GFP Control Fishell 99130 pAAV-mDlx-NLS-mRuby2....AX with GRE or transgenic animal Oligodendrocytes - AAV-PHP.eB with GRE or transgenic animal Brain vascular....AX with GRE or transgenic animal L5 & Inhibitory - AAV-PHP.eB with GRE or transgenic animal L2/3 & L4...AAV.CAP-B10 with GRE or transgenic animal Excitatory - AAV.PHP.N with GRE or transgenic animal For PNS applications... pAAV-S5E2-GFP-fGFP E2 regulatory element GFP, membrane-targeted GFP Control Dimidschstein 192552 pAAV-CAG... viral prep format. Please note this does not guarantee viral vector packaging service, but lets us know...

...packaging transfection. In my lab we regularly use the ratios below: 1 Transfer Plasmid : 1 pHelper: 1 RepCap... plan to transfect: 1 to 10 Fill in experimental details on the spreadsheet: Name your viral prep...calculator in the comments section below. References 1. Choi, Vivian W., et al. "Production of Recombinant...prevent future headaches. To help you plan PEI transfections for AAV packaging, considering using this PEI...Calculator. AAV packaging typically requires transfecting three plasmids at specific molar ratios. To ...PEI calculator for AAV packaging Use the PEI transfection calculator for AAV packaging Select the plasmid...RepCap OR 3 Transfer Plasmid : 5 pHelper: 2 RepCap Select what type of tissue culture dish you will be ...

...integration. Introduction to Viral Vectors Figure 1: Wild-type virus propagation and use of viral plasmids...packaging capacity, high transduction efficiency, and high but transient transgene expression.They can induce...130 nm Dividing Yes Moderate AAV Transient or Stable* Single-stranded linear DNA ∼4.5 kb 18–26 nm Dividing.../Non-dividing No* Very Low Adenoviral Transient Double-stranded linear DNA 5–36 kb 105 nm Dividing/Non-dividing...of viral vectors and find viral production and transfer plasmids for a wide array of research purposes...are versatile tools because of their ability to transduce dividing and non-dividing cells. These virusal...immunogencity. These viral vectors can direct long-term transgene expression, but generally don't integrate into...

...formulation buffer is phosphate-buffered saline containing 1 mM sodium azide. This buffer is not compatible for...extracted from cells, separated by SDS-PAGE, transferred to a membrane, and labeled with the antibody and a species-specific...expression vectors, human embryonic kidney cells are transfected with the plasmid(s) encoding the heavy and light...anti-GFP antibody), the target antigen is first transiently expressed in mammalian cells. To learn which ... The fluorescence pattern is compared to an untransfected control. Immunohistochemistry (IHC) Tissue sections...antibody. Protein expression is compared to an untransfected control and the protein size is confirmed. Residual...contain low levels of co-purified plasmid from transfection. This residual plasmid is extracted and amplified...

...cell surface proteins. J Am Chem Soc. 127(37):12770-1. PMID:16159249...with a recognizable sub-cellular localization (membrane, nucleus, etc.) are also available Non-toxic to... is based on the human O6-alkylguanine-DNA-alkyltransferase (hAGT), a DNA repair protein, and is suitable...Vector SNAP-tag Empty backbone for stable and transient mammalian expression 101133 pSNAP-CaaX Control...Control Plasmid SNAP-tag Control for endomembrane (CaaX box) localization 101129 pSNAPf-Cox8A Control Plasmid...Vector CLIP-tag Empty backbone for stable and transient mammalian expression 101134 pCLIPf-H2B Control...pMCP-tag (m) Vector ACP/MCP-tag Empty backbone for transient mammalian expression 101126 pACP-tag (m)- 2 Vector...

... your account. We will notify you by email within 1 business day regarding your account verification. ... Material Transfer Agreements Most Addgene orders must be accompanied by a material transfer agreement...industry scientists. Please note this does not guarantee availability but lets us know what plasmids would...agreement (MTA), including transfers to for-profit entities. The MTA is a legal agreement between the recipient... the depositing institution(s). It governs the transfer of biological materials and intellectual property...Please see the Sample Agreements on our Technology Transfer Information page for more information. To learn...

... find a particular viral spike protein (MERS, CoV-1, CoV-2), modification, or variant. ID Plasmid Description...provirus and the overall strength of viral transcriptional response in these cells. Warner Greene 105621...

...available as a substrate for another round. Figure 1: Overview of prime editing. Created with BioRender.com...incorporated into the DNA by reverse transcription. The edited DNA strand displaces the unedited 5’ flap and...replace” gene editing method in which a reverse transcriptase (RT) is fused to the C terminus of Cas9 H840A...the target sequence where it nicks the non-target strand and generates a 3’ flap. The 3’ flap binds to the...Moloney murine leukemia virus (M-MLV) reverse transcriptase. Engineering of the PE enzyme and related components...including exploring new sources for the reverse transcriptase. Many PE enzymes and related tools are available...

...type of choice in vitro is known as reprogramming [1]. The process can be divided into two stages: Dedifferentiation...//www.linkedin.com/in/kmukherjeephd/.  References 1. Hochedlinger, K. and R. Jaenisch, Nuclear reprogramming...from human peripheral blood. Cell Stem Cell, 2010. 7(1): p. 15-9. PubMed PMID: 20621044. PubMed Central PMCID...induced pluripotent stem cells. Hepatology, 2010. 51(1): p. 297-305. PubMed PMID: 19998274. PubMed Central... pluripotent stem cells. Nat Biotechnol, 2014. 32(1): p. 84-91. PubMed PMID: 24291815. PubMed Central ...keratinocyte lineage. Methods Mol Biol, 2014. 1195: p. 1-12. PubMed PMID: 24510784. PubMed Central PMCID: PMC4096605...induced pluripotent stem cells. PLoS One, 2010. 5(1): p. e8763. PubMed PMID: 20098701. 31. Tang, Z.H., et ...

...serotypes. ID Plasmid Capsid Description PI 112862 pAAV2/1 AAV1 AAV packaging plasmid, expressing Rep2 and Cap1...use three plasmids: A transfer plasmid (or cis plasmid) containing the transgene or genetic elements of...service gives you even more options. AAV Transfer Plasmids Transfer plasmids encode the genetic information...Browse AAV transfer, packaging, and helper plasmids in Addgene's repository. Viral...Plasmids Adeno-Associated Viral (AAV) Plasmids AAV Transfer Plasmids AAV Packaging Plasmids AAV Helper Plasmids...precise delivery of genetic material across a wide range of tissue types with minimal immune response in ...This collection provides a curated selection of transfer, packaging, and helper plasmids that can be used...

...details on how they work!) Figure 1: Base transitions are edits between the purine bases (adenine... inducing targeted DNA transversions in human cells. Nature Biotechnology, 39(1), 41–46. https://doi.org...R-loop structure. In order to bind DNA, Cas9 requires (1) a guide RNA, (2) a complementary DNA sequence, and...given the pithier name CGBE1, for “C → G Base Editor 1,” and had a C → G editing efficiency of up to 68%... the CGBE: DNA polymerase D2 (POLD2), exonuclease 1 (EXO1), and RNA binding motif protein X-linked (RBMX...excision repair proteins. Nature Communications, 12(1), 1384. https://doi.org/10.1038/s41467-021-21559-9...machine learning methods. Nature Communications, 12(1), 4902. https://doi.org/10.1038/s41467-021-25217-y...

...component involved in GTPase signaling activity. Figure 1: Locations of ARF superfamily members. Figure from...regulators of membrane traffic, they also play critical roles at cilia, centrosomes, the plasma membrane, lipid...2016). Tracing the archaeal origins of Eukaryotic Membrane-Trafficking System building blocks. Molecular ...., Dacks, J. B., Lambright, D. G., Lee, F. S., Randazzo, P. A., Santy, L. C., Schürmann, A., Wilhelmi,...

...activity in single cells using translocation reporters. Nature protocols, 13 1, 155-169. PMID: 29266096. Find...Kinases: they regulate many proteins, with ~1/3 of human proteins predicted to be phosphorylated on at...which have previously been characterized. See Box 1 in Kudo et al. for more details on the design of the...phosphatase-accessible. Kinase Translocation Reporters (KTRs) Kinase Translocation Reporters (KTRs) are genetically...particularly important for regulating signal transduction and measuring kinase activity at the single-...provides an alternative tool with their Kinase Translocation Reporters (KTRs) whose cellular localization...activity is enhanced, leading to cytoplasmic translocation of the reporter. As a result, the KTR is shuttled...

...Table 1 has a detailed list of the bacterial species these plasmids are compatible with.  Table 1: Non-...Figure 1: Idealized examples of a narrow host range plasmid with only one ori and a broad host range plasmid...Improved broad-host-range plasmids for DNA cloning in Gram-negative bacteria. Gene, 70(1), 191–197. https... N.J.), 765, 327–343. https://doi.org/10.1007/978-1-61779-197-0_19  Meyer, R. (2009). Replication and ...?  Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Tranfection...define plasmid host range, identify a few of the features that confer broad host range in plasmids in nature...features to create broad host range recombinant vectors.  A plasmid’s host range is determined by the different...

...138920210790886871 Pray, L. (2008) Transposons: The jumping genes. Nature Education 1(1):204. https://www.nature.com...reverse transcriptase activities.    Figure 1: Overview of retrotransposon transposition. Retrotransposons...are also known as retrotransposons. They transpose by a ‘copy-paste’ mechanism (Fig. 1). They first copy...their host’s genome.   There are two types of Class 1 TEs: those with long terminal repeats (LTRs) and those...CRISPR-Cas system. This system has four major components: 1) a CRISPR RNA (crRNA), 2) four proteins (TniQ, Cas8...express a transposase or reverse transcriptase so they can transpose. Both class I and II transposons can be...eukaryotic genomes.  Components of a transposon system Transposon: The transposon is the DNA sequence that moves...

...coming closer to this DNA targeting ideal. Figure 1: (A) Representation of a zinc finger (ZF) nuclease..., that will probably never happen. Yet with transcription activator-like (TAL) effectors, scientists in...sequence-specific TAL effectors for modulating mammalian transcription. Nat Biotechnol. 2011 Feb;29(2):149-53. PMID...

... compounds with various antimicrobial activities (1). She has developed a fast endophyte screening system...us coordinate and promote the awards. References 1. Martinez-Klimova, Elena et al. “Endophytes as sources...antibiotic” Biochemical Pharmacology 2017 Jun 15;134:1-17. PubMed PMID: 27984002. 2. Bilan, Dmitry S. et ...Addgene-Seeding Labs Plasmid Grant Check Out Previous Addgene-Seeding Labs Plasmid Grant Awardees Listen to Our...Labs and Addgene. The Addgene-Seeding Labs plasmid grant is a collaborative endeavor providing plasmids to...who were awarded the Addgene-Seeding Labs Plasmid grant this year. These scientists work on projects spanning...and to increase the potential for this drought tolerant plant, Dr. Robertson aims to make use of the CRISPR...

...to determine purity of the AAV preparation. Figure 1: Silver staining of purified and non-purified AAV ...of Pennsylvania Vector Core. Transfections are performed using the transfer plasmid, a plasmid encoding...cell cultures are inspected for sterility. Transducibility Some viral vectors are tested in vitro and ...section for images). Figure 2: AAV Pro cells were transduced with either pAAV-Ef1a-mCherry-IRES-Cre (55634...filtration through a 0.45 µm polyethersulfone (PES) membrane. The preparations are then aliquoted, frozen, ...opens in a new window) ). Briefly, 293T cells are transduced with serial dilutions of a lentiviral vector,... taken off-line and discarded. Confirmation of Transfer Plasmid Addgene uses a rigorous barcode matching...

...each reporter or check out table 1 for a summary. HRP HRP tagged RANbodies don’t require a secondary antibody... Table 1: Summary of the Key Features of HRP, Spaghetti Monster, and Chicken IgY Domain RANbody Reporters...NIH’s Developmental Studies Hybridoma Bank (DSHB). 1 ml of antibody-containing hybridoma supernatant is... generate and costly to share. What is a RANbody? A RANbody is a fusion between a reporter and a nanobody...nanobody sequence could be used to make a RANbody. RANbodies are expressed in 293T cells and are secreted...smaller size of RANbodies compared to antibodies. Staining with multiple HRP-RANbodies is indistinguishable...HRP-conjugated RANbody can be used at a time. Samples can be sequentially stained with two HRP RANbodies if the...

...course much more difficult than in cell culture, but Ran et al. have shown that CRISPR and AAV can be used...Thakore PI, Moreb EA, Rivera RM, Madhavan S, Pan X, Ran FA, Yan WX, Asokan A, Zhang F, Duan D, Gersbach CA...WL, Widrick JJ, Yan WX, Maesner C, Wu EY, Xiao R, Ran FA, Cong L, Zhang F, Vandenberghe LH, Church GM, ...: 25692716. PubMed Central PMCID: PMC4335351. 9. Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz...X-linked recessive disorder affecting approximately 1 in 5000 males. DMD is caused primarily by frameshift... applicable to 80% of DMD patients. Figure 1: Exon skipping strategies for Duchenne muscular dystrophy...needed. First, CRISPR delivery must be optimized to: 1. Reach a high percentage of muscle cells throughout...

...obstacles when working with virus: 1. Viral vectors undergo DNA rearrangements. Depending on the composition...approximately 20–50% of the packaging cells after 1–3 days of selection. A lower percentage of surviving...up-to-date tips for your virus experiments. References 1. Anthony, Simon J., et al. "A strategy to estimate...may undergo genomic rearrangements during replication. To avoid genomic rearrangements, try amplifying the...immediately prior to their transduction has been shown to increase AAV transduction efficiency by approximately...immediately following transfection. In other words, when exchanging the medium after transfecting cells, replace...Fibronectin is another membrane-interacting protein that can be used to enhance transduction efficiency in cells...

...if you obtained a transformant with a single insertion because it will segregate 3:1 in the T2 generation...lineage and has made countless transgenic plants in the process.     References 1. Emami, S., Yee, M. and Dinneny...Methods. BioMed Central, 2(1), p. 19. doi: 10.1186/1746-4811-2-19. PubMed PMID: 17087829. PubMed Central... which is ease of transformation. There are many motivations to generate transgenic Arabidopsis, from ...from studying transcriptional and translational dynamics of genes and proteins in living plants, to complementing... dip transformation method. The general steps for this method include: Cloning and transforming a plasmid...integrates transfer DNA (tDNA) into the genomes of the plants it attacks Growing the transformed agrobacterium...

...focus on the AAV transfer plasmid and take a look at each of its parts. Figure 1: Parts of an AAV...human synapsin 1 promoter is neuronal specific (Haery et al., 2019). Most AAV transfer plasmids have a...repeats (ITRs) are what makes an AAV transfer plasmid an AAV transfer plasmid. ITR sequences are 145 bases...read through this challenging sequence. Transgene The transgene is the gene that will be delivered by the...control transgene expression. Lox sites are directional 34 bp sequences that can flank the transgene and ...(WHP) Posttranscriptional Regulatory Element (WPRE) forms a tertiary structure when transcribed which ... nuclear export of RNA and RNA translation, and promotes RNA transcript longevity. The polyA is located...

...Check out Addgene's New Viral Service! References 1. Hay, R.J., Macy, M.L. & Chen, T.R. “Mycoplasma infection...contaminations.” Methods in Molecular Biology 946 (2013). 1-13. Pubmed PMID: 23179822. 5. Segura, M.M., Kamen,...Production, concentration and titration of pseudotyped HIV-1-based lentiviral vectors."Nature protocols 4.4 (2009...may not translate well to other cell types. To overcome this issue, try transducing with a range of dilutions...DEAE-dextran can improve transduction efficiency. In addition, this study found that transduction efficiency...passage number of a culture the lower the transfection and transduction efficiencies. Cultures should only be...others opt for a single harvest after transfecting with the transfer and packaging plasmids. A benefit of...