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We narrowed to 114 results for: N-pac

Showing: 1 - 40 of 114 results

  1. Viral Vectors 101: Parts of the AAV Packaging Plasmid

    Type
    Blog Post
    Published
    Dec. 3, 2020, 2:15 p.m.
    ...C-terminal regions of the Rep proteins. While the unique N-terminal domain of Rep78 and Rep68 contains a DNA-...experiment:  the packaging plasmid which contains the AAV structural and packaging genes, the adenoviral...on the packaging plasmid and take a look at each of its parts. Key elements of the AAV packaging plasmid...trans, freeing up space in the viral genome so that a gene of interest can be packaged into the virus instead...plasmid The packaging plasmid aka the RepCap plasmid encodes two genes: the replication (rep) gene and the...are required for viral genome replication and packaging: Rep78, Rep68, Rep52, and Rep40. Rep gene expression...Rep proteins from the AAV2 serotype are used to package vector genomes flanked by AAV2 ITRs into a capsid...

  2. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    Published
    Oct. 24, 2017, 1:54 p.m.
    ...represent the views of Addgene. Boston, MA, USA 42.3601°N 71.0589°W – Alina Chan The job of scientists to observe...artificial chromosomes. Montreal, QC, Canada 45.5017°N 73.5673°W – Vanessa Sung Hundreds of scientists and...on twitter @sung_vanessa. Albany, NY, USA 42.6526°N 73.7562°W – Nicholas Thompson Over the last few years...twitter at @thompn4. New York City, NY, USA 40.7128°N 74.0059°W – Deanna McLafferty I’m a New Yorker, so...assistant and activist. Washington, DC, USA 38.9072°N 77.0369°W – Mark Lucera I am a passionate scientist...twitter at @Mark_Lucera. Ann Arbor, MI, USA 42.2808°N 83.7430°W – Alexandra Greenbaum Science and evidence-based...doesn't tweet enough.  Nashville, TN, USA 36.1627°N 86.7816°W- Jennifer Stancill I am a research biologist...

  3. The Advances Behind The World's First Personalized CRISPR Treatment

    Type
    Blog Post
    Published
    Sept. 16, 2025, 1:15 p.m.
    ...10.1038/nature24644  Gaudelli, N. M., Lam, D. K., Rees, H. A., Solá-Esteves, N. M., Barrera, L. A., Born, ...Silverstein, R. A., Hille, L. T., Ogul, A. N., Robinson-Garvin, N. A., Small, J. C., McCague, S., Burke, ...10.1126/science.1231143  Gaudelli, N. M., Komor, A. C., Rees, H. A., Packer, M. S., Badran, A. H., Bryson,...Silverstein, R. A., Kim, N., Kroell, A.-S., Walton, R. T., Delano, J., Butcher, R. M., Pacesa, M., Smith, B. K...wild-type enzyme requires a PAM sequence of NGG (where N represents any base), their engineered variants are...Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P. D., Wu, X., Jiang, W., Marraffini, L. A....-y  Walton, R. T., Christie, K. A., Whittaker, M. N., & Kleinstiver, B. P. (2020). Unconstrained genome...

  4. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    Published
    Oct. 12, 2023, 1:15 p.m.
    ... A. R., Shin, N., Kim, Y., Toong, N., Kaplow, I. M., Wirthlin, M., Zhang, X., Phan, B. N., Fox, G. A.,...s41467-023-38582-7 Chuapoco, M. R., Flytzanis, N. C., Goeden, N., Christopher Octeau, J., Roxas, K. M., Chan.../10.1038/nbt.3440 Goertsen, D., Flytzanis, N. C., Goeden, N., Chuapoco, M. R., Cummins, A., Chen, Y., ...10.1038/s41593-021-00969-4 Goertsen, D., Goeden, N., Flytzanis, N. C., & Gradinaru, V. (2022). Targeting the...Radaelli, C., Gore, B. B., Weed, N., Omstead, V., Bishaw, Y., Shapovalova, N. V., Martinez, R. A., Fong, O... AAV9.452sub.LUNG1 Lung: ATII cells Mice N/A Goertsen et al., 2022 AAV.CAP-B22 CNS: neurons...Rats, marmosets, green monkeys, rhesus macaques N/A Chuapoco et al., 2023 AAV-F CNS: neurons...

  5. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    Published
    Sept. 9, 2014, 8:20 p.m.
    ... G D V E E N P G P P2A: (GSG) A T N F S L L K Q A G D V E E N P G P E2A: (GSG) Q C T N Y A L L K L... L A G D V E S N P G P F2A: (GSG) V K Q T L N F D L L K L A G D V E S N P G P * (GSG) residues can...may take up precious space in viral transfer vectors with limited packaging capacity. Additionally, it may...particularly when only a portion of the plasmid is packaged for viral delivery, or the relative expression...

  6. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    Published
    Feb. 13, 2025, 2:15 p.m.
    ...10.1038/nature24644 Gaudelli, N. M., Lam, D. K., Rees, H. A., Solá-Esteves, N. M., Barrera, L. A., Born, ..., Schwaegerle, N. D., Peraro, L., Young, L., Lee, S.-J., Ciaramella, G., & Gaudelli, N. M. (2023). Improved...Neugebauer, M. E., Hsu, A., Arbab, M., Krasnow, N. A., McElroy, A. N., Pandey, S., Doman, J. L., Huang, T. P....10.1038/s41587-022-01532-7 Gaudelli, N. M., Komor, A. C., Rees, H. A., Packer, M. S., Badran, A. H., Bryson,...nature17946 Komor, A. C., Zhao, K. T., Packer, M. S., Gaudelli, N. M., Waterbury, A. L., Koblan, L. W.,...byproducts likely resulted from excision by uracil N-glycosylase (UNG) during base excision repair. Adding..., the team fused bacteriophage protein Gam to the N-terminus of BE4 (Komor et al., 2017). Gam binds the...

  7. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    Published
    April 1, 2025, 1:15 p.m.
    ... Uracil DNA N-Glycosylase Removes uracil from DNA, creating an abasic site. MPG / AAG N-methylpurine... site using either fused or endogenous Uracil DNA N-glycosylase (UNG) enzymes. Various other proteins ...mammals also have an enzyme to remove inosine from DNA: N-methylpurine DNA glycosylase (MPG), also known as ...language models-assisted optimization of a uracil-N-glycosylase variant enables programmable T-to-G and...doi.org/10.1016/j.molcel.2024.01.021 Tong, H., Liu, N., Wei, Y., Zhou, Y., Li, Y., Wu, D., Jin, M., Cui,...10.1093/nsr/nwad143 Tong, H., Wang, H., Wang, X., Liu, N., Li, G., Wu, D., Li, Y., Jin, M., Li, H., Wei, Y....s41467-024-49343-5 Tong, H., Wang, X., Liu, Y., Liu, N., Li, Y., Luo, J., Ma, Q., Wu, D., Li, J., Xu, C.,...

  8. "Hall of Fame" AAV Enhancers from the Allen Institute for Brain Science

    Type
    Blog Post
    Published
    Sept. 9, 2025, 1:15 p.m.
    ..., Johansen, N. J., Hooper, M., Omstead, V., Vargas, S., Lerma, M. N., Taskin, N., Weed, N., Laird, W. ...Radaelli, C., Gore, B. B., Weed, N., Omstead, V., Bishaw, Y., Shapovalova, N. V., Martinez, R. A., Fong, O... Kalmbach, B., Lenz, G. H., Morin, E., Nguyen, T. N., Garren, E., Bendrick, J. L., Kim, T. K., Zhou, T...help propel research forward and have the biggest impact. We should know — Addgene has built its foundation...color":"#0052a9","css":"#0052a9","hex":"#0052a9","opacity":100,"rgb":"rgb(0, 82, 169)","rgba":"rgba(0, 82..."css":"rgba(0, 82, 169, 75%)","hex":"#0052a9","opacity":75,"rgb":"rgb(0, 82, 169)","rgba":"rgba(0, 82,...color":"#ffffff","css":"#ffffff","hex":"#ffffff","opacity":100,"rgb":"rgb(255, 255, 255)","rgba":"rgba(255...

  9. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...CIBN, where CIBN is the N-terminal fragment of CIB1 and it was fused to both the N- and C-termini of dCas9...transcription of the N-terminus is light-dependent (Yu et al., 2020). To express the N-Cas9, Haifeng Ye’s...activators p65 and VP64 which in turn transcribes N-Cas9. The N- and C-Cas9 fragments then dimerize to form ...photoreceptor, Vivid (VVD, N. crassa) (Kawano et al., 2015). Nicknamed paCas9-1 and consisting of the fusion...transcription Nihongaki et al., 2015 paCas9 Magnet proteins from N. crassa Split Cas9, Split Cas9 nickase...split” nuclease - the authors fragmented Cas9 into N-terminal (residues 2-713, N713) and C-terminal (residues...https://doi.org/10.1093/nar/gkw930 Yu Y, Wu X, Guan N, Shao J, Li H, Chen Y, Ping Y, Li D, Ye H (2020) Engineering...

  10. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    Published
    Jan. 16, 2025, 2:15 p.m.
    ...Capsid Primary receptor Coreceptor AAV1 α2,3-/α2,6 N-linked sialic acids1 AAV receptor (AAVR)2   AAV2...HGFR)3 Laminin receptor (LR)3 CD93 AAV5 2,3-N-linked sialic acid4 AAV receptor (AAVR)2 Platelet-derived...factor receptor (PDGFR) α and β3 AAV6 α2,3-/α2,6 N-linked sialic acids1 Heparan sulfate proteoglycan ...D (CPD), also called AAVR25   AAV9 Terminal N-linked galactose3 AAV receptor (AAVR)2 Laminin...doi.org/10.1128/jvi.75.14.6615-6624.2001 Han, Z., Luo, N., Ma, W., Liu, X., Cai, Y., Kou, J., Wang, J., Li,...785. https://doi.org/10.3390/cells12050785 Kaludov, N., Brown, K. E., Walters, R. W., Zabner, J., & Chiorini...//doi.org/10.1089/hum.2019.173 Pillay, S., Meyer, N. L., Puschnik, A. S., Davulcu, O., Diep, J., Ishikawa...

  11. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    Published
    Sept. 16, 2020, 1 p.m.
    ...  Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini LA, Zhang F (2013.../nature09523  Ibraheim R, Song C-Q, Mir A, Amrani N, Xue W, Sontheimer EJ (2018) All-in-one adeno-associated...doi.org/10.1038/ncomms14500  Levy JM, Yeh W-H, Pendse N, Davis JR, Hennessey E, Butcher R, Koblan LW, Comander...doi.org/10.1038/s41551-019-0501-5  Liu J-J, Orlova N, Oakes BL, Ma E, Spinner HB, Baney KLM, Chuck J, Tan..., Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O (2014) Crystal Structure...08224-4  Swiech L, Heidenreich M, Banerjee A, Habib N, Li Y, Trombetta J, Sur M, Zhang F (2014) In vivo ...into an AAV vector is challenging due to its packaging capacity of AAV (~4.7 kb). While this approach has...

  12. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    Published
    April 9, 2019, 1:13 p.m.
    ...biosensors (yellow cameleon 3.60, EPAC, and Galphai), the C-terminus and N-terminus of the donor fluorescent...increase the FRET efficiency. Creating fusions at the N- and C- termini of your protein of interest The most...straightforward fusion protein is made by connecting the N- or C-terminus of the fluorescent protein and protein...to describe the different parts of the fusion from N- to C-terminus. For instance, Lifeact-mTurquoise2 ...We have encountered several proteins for which the N- and C-terminus of the POI are necessary for its native...fluorescent protein into the POI can work well since the N- and C-terminal residues of the fluorescent protein.... Klarenbeek, Jeffrey, et al. "Fourth-generation epac-based FRET sensors for cAMP feature exceptional ...

  13. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    Published
    Sept. 6, 2022, 1:15 p.m.
    ...doi.org/10.1089/hgtb.2019.105 Rumachik, N. G., Malaker, S. A., Poweleit, N., Maynard, L. H., Adams, C. M., Leib...detailed methods and image descriptions, a minimum n = 2, and well labeled high- and low-magnification ...Connor, D. M., Lutomski, C., Jarrold, M. F., Boulis, N. M., & Donsante, A. (2019). Lot-to-Lot Variation in...Stamnes, S., Holt, K., Sinn, P., May, A. P., & Paulk, N. K. (2020). Methods Matter: Standard Production Platforms...much the spread of the virus to adjacent areas impacted the experiments we wanted to perform. It required...injection titers (1,2), and cell types used for packaging viral particles (6), can influence vector performance...

  14. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    Published
    March 25, 2025, 1:15 p.m.
    .../advs.202202957 Yuan, T., Yan, N., Fei, T., Zheng, J., Meng, J., Li, N., Liu, J., Zhang, H., Xie, L., ...and instead introduced its opposite, a uracil DNA N-glycosylase (UNG) from E. coli. The resulting construct..., improved editing efficiencies when fused to the N terminus of a GBE construct. This effect improved ... / apyrimidinic (AP) site. UNG / UDG Uracil DNA N-Glycosylase Removes uracil from DNA, creating an abasic...P. D., Prekop, H.-T., Chew, Y. T., Manivannan, S. N., & Chew, W. L. (2021). Programmable C:G to G:C genome...S., Liu, L. D., Zhang, S., Zhang, Y., Wo, Y., Li, N., Wu, S., Gui, T., Wang, B., Luo, Y., Cai, Y., Liu... (2) a complementary DNA sequence, and (3) a protospacer-adjacent motif (PAM) on the opposite strand. ...

  15. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    Published
    Oct. 27, 2015, 3 p.m.
    ...FPs that encode N-glycosylation consensus sequences are modified with the addition of N-glycans [9]. Both...cloning strategy and decide whether you will create an N- or C-terminal fusion based on your experimental needs...were re-engineered by mutating cysteine residues and N-glycosylation consensus sequences in popular FPs. ...investigators realize too late that they have missed the impact of dark, non-fluorescent, and misfolded FP-fusions...quantitative imaging experiments or even negatively impact cells (see Figure 1). When behaving properly, ... the function of resident ER chaperones and may impact cellular function or possibly viability. FP misfolding.... It remains unclear whether the FP misfolding impacts fusion protein function and this could lead to ...

  16. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    Published
    July 11, 2023, 1:15 p.m.
    ...10.1016/j.neuron.2016.01.040 Saloman, J. L., Scheff, N. N., Snyder, L. M., Ross, S. E., Davis, B. M., & Gold...muscarinic receptor derived DREADDs has been clozapine-N-oxide (CNO). However, concerns have been raised over...Pharmacokinetic and pharmacodynamic actions of clozapine-N-oxide, clozapine, and compound 21 in DREADD-based ...allows you to study signaling pathways and the impacts of neural circuits on behavior. Figure 1:...channels. When working with PSAM receptors, your impact on the cell is determined by what IPD your PSAM...

  18. Plasmids 101: Cre-lox

    Type
    Blog Post
    Published
    Jan. 13, 2015, 3:47 p.m.
    ...created, which contain a unique asymmetric spacer "NNNTANNN", where "N" indicates which bases may vary from .... I. Recombination between loxP sites. Sternberg, N. and Hamilton, D. 1981. PubMed PMID: 6276558. 2. Site-specific...recombinase of bacteriophage P1. Sauer, B. and Henderson, N. 1988. PubMed PMID: 2839833. PubMed Central PMCID:...repeats separated by an 8-bp long asymmetric core spacer sequence. The asymmetry in the core sequence gives... double-strand DNA break occurs within the core spacer of each loxP site and the two strands are ligated...

  19. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    Published
    Oct. 19, 2023, 1:15 p.m.
    ... S., Lee, W., Walsh, N., Iozza, K., Huang, N., Preston, G., Drouin, L. M., Jia, N., Deng, J., Hebben, ...expression at varying capacities. AAV vectors containing the AAV2 ITRs can be cross-packaged with capsids from...replication and are required for replication, packaging, and vector persistence. They work in conjunction...in the same DNA molecule as the gene that they package. In a transfer plasmid, the transgene is flanked...deleting the B-B' or C-C' arms can reduce the packaging efficiency but increase transgene expression (...Terminal Repeats (ITRs) on the Productivity, Genome Packaging, and Potency of AAV Vectors. Microorganisms, 12...Bonnin, D., Merten, O.-W., & Galibert, L. (2017). Impact of Inverted Terminal Repeat Integrity on rAAV8 ...

  20. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...dimerization. Splitting wild-type Cas9 into n-terminal (Cas9(N)-2xNLS) and c-terminal (Cas9(C)-2xNLS) fragments...2xNLS) and the n-terminal cas9 fragment with FKBP rapamycin binding domain of mTor (Cas9(N)-FRB-NES) resulting...Split Cas9 system The Cas9 protein is composed of an n-terminal DNA recognition domain and a mostly c-terminal...genome editing. Without rapamycin treatment, the Cas9(N)-FRB-NES fragment is actively shuttled out of the ...export sequence. Treatment with rapamycin induces Cas9(N)-FRB-NES and Cas9(C)-FKBP-2xNLS dimerization and net...activator fragments (dCas9(C)-FKBP-2xNLS-VP64 and dCas9(N)-FRB-NES). This system provides users greater temporal...for constructing fusion proteins with customizable N- and C-terminal modules   Thanks to the lab of Tomás...

  21. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ...s41587-021-01133-w Pandey, S., Gao, X. D., Krasnow, N. A., McElroy, A., Tao, Y. A., Duby, J. E., Steinbeck...efficacy.   Find CHARM plasmids here!   Neumann, E. N., Bertozzi, T. M., Wu, E., Serack, F., Harvey, J. ...., Pirtle, C. P., Coffey, A., Howard, M., Kamath, N., … & Weissman, J. S. (2024). Brainwide silencing .... Z., Replogle, J. M., Adriaens, C., Ramadoss, G. N., Shi, Q., Hung, K. L., Samelson, A. J., … & Weissman...evolved optimizations for prime editing CHARM: A compact epigenetic silencer Recombinant antibodies for ...assembly of biotinylated SpyTag-SpyCatcher linkers and spacer proteins ending with a GFP nanobody (Figure 1)....determination. A) Schematic of workflow. B) Detail of spacer peptides (3HB: 11-nm 3-helix bundle; SAH: 60-nm...

  22. Hot Plasmids: Spring 2025

    Type
    Blog Post
    Published
    June 3, 2025, 1:15 p.m.
    ...Sanchez, A. D., Udeshi, N. D., Svinkina, T., Carr, S. A., Feldman, J. L., Perrimon, N., & Ting, A. Y. (2018...Flexible tagging: Choose untagged or add a versatile N- or C-terminal HA tag. Selectable and stable: A puromycin... Figure 2: Addgene cloned EGFP into pAG Lenti CMV N-HA Puro (Addgene #236079) to create pAG Lenti CMV ...needed to replicate inside a host cell but with space to insert your genetic cargo of choice. After hearing...taken them for a test drive! We validated viral packaging and strong expression after inserting EGFP and...

  23. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ...acrIIA3   Listeria monocytogenes  pCSW65 N/A (Toxic)   None N/A   acrIIA4 Listeria monocytogenes  pCSW21...Gateway-Compatible Toolbox Podcast Segment The impact of KLF4 N-terminal variants on iPSC generation Deriving...fluorophores with nonoverlapping emission spectra (N- and C-terminal tags for mTagBFP, TagRFPt, EGFP, mVenus...discrepancies, which they traced to a 9 amino acid N-terminal variation in the Klf4 isoform (KLF4s or KLF4L...pJH376 Yes   acrIIA3 Streptococcus pyogenes  pCSW24  N/A (Toxic) pJH375  No    ... expression levels; this altered stoichiometry impacted reprogramming and global gene expression patterns...

  24. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    Published
    July 16, 2024, 1:15 p.m.
    ...hgtb.2011.034 Benyamini, B., Esbin, M. N., Whitney, O., Walther, N., & Maurer, A. C. (2023). Transgene Expression...  References  Aurnhammer, C., Haase, M., Muether, N., Hausl, M., Rauschhuber, C., Huber, I., Nitschko,...adenoviral helper genes are supplied on two different packaging plasmids: the pHelper plasmid containing Ad E2A... your infectious titer, since it will have more impact on your experimental outcome and planning.  Both... of AAV-2 particles via a novel capsid ELISA: Packaging of genomes can limit production of recombinant...resources on the Addgene blog Parts of the AAV Packaging PlasmidParts of the AAV Transfer PlasmidAAV Variables...

  25. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...Terminus 79629 (N) and 79628 (C) Cv (capsid) N and C 79633 (N) and 79641 (C) NS1 (var: W98G) N and C 79635...NS2B C 79636 NS4A N 79640 NS4B C 79639 NS5 C 79632 (N) and 79631 (C) PrM N and C   Detecting...effects. The library was cloned into the Liu lab’s own pAc-sgRNA-Cas9 insect expression backbone, which expresses.... The Roux lab has deposited HA tagged BioID2 for N-terminal fusions and Myc tagged BioID2 for C-terminal... (< 100 bp) and large (up to 120 kb) deletions in N. benthamiana and Arabidopsis. The authors found that...be found here). To use the library, the authors packaged it into lentivirus and used the resulting lentiviral...large size of dCas9-VPR limits one’s ability to package it into lentivirus and later create stable cell...

  26. Plasmids 101: A Brief History of Plasmids and an Improved eBook!

    Type
    Blog Post
    Published
    Oct. 29, 2015, 2:30 p.m.
    ...Personal Perspective: Plasmid (1952-1997) Cohen, Stanley N. “DNA Cloning: A Personal View after 40 Years.”Proceedings... PubMed Central PMCID: PMC2134509. Cohen, Stanley N. et al. “Construction of Biologically Functional Bacterial...plasmids and new cloning vectors such as pBR322, pACYC, and pUC were developed to provide higher copy number... discovery in the 1950s, plasmids have greatly impacted many areas of biological research and have been...

  27. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...inevitable. Most FP fusions for FRET studies are at the N- or C-terminus of your target protein, but even these... Royal Society of Chemistry. 3. Ermolenko, Dmitri N., et al. "Observation of intersubunit movement of ...272. PubMed PMID: 9657724. 6. Kapanidis, Achillefs N., Yon W. Ebright, and Richard H. Ebright. "Site-specific...fluorescent labeling with (Ni2+: nitrilotriacetic acid) n-fluorochrome conjugates." Journal of the American ... a bit like a Fedex delivery. Your fluorophore package needs to go to the proper mailbox (i.e., a binding...that can nonspecifically soak up your fluorophore package. One important concept in FRET experimental design...synthetase for p-benzoyl-l-phenylalanine E. coli pAcBac1.tR4-MbPyl pyrrolysyl-tRNA synthetase Mammalian ...

  28. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    Published
    Sept. 24, 2024, 1:15 p.m.
    ...Polali, S., Anderson, E. D., Bell, A. M., Tzouanas, C. N., Avants, B. W., & Robinson, J. T. (2019). Magnetic...andrewgyork.github.io/gfp_magnetofluorescence. Karki, N., Vergish, S., & Zoltowski, B. D. (2021). Cryptochromes...https://doi.org/10.1038/nchem.2447.  Lee, H., Yang, N., & Cohen, A. E. (2011). Mapping Nanomagnetic Fields...-up studies. “We don’t have a vision of passive impact,” he says, noting that his team regularly shares...

  29. An Introduction to Adenovirus

    Type
    Blog Post
    Published
    Oct. 3, 2023, 1:15 p.m.
    ...10400791. Kafri, T., Morgan, D., Krahl, T., Sarvetnick, N., Sherman, L., & Verma, I. (1998). Cellular immune...Foresythe, A., Huh, H. J., Bazinet, A. L., Bergman, N. H., Bull, R. L., Garcia, K. Y., Goodrich, J. S., ....1038/s41586-023-05949-1. PMID: 36996871. Tatsis, N., Fitzgerald, J. C., Reyes-Sandoval, A., Harris-McCoy..., including the E1 region). The space in the viral particle’s capacity that is made available by this ...genome, and adenoviral vectors all have a genomic capacity that is much higher than other viral vectors (...

  30. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    Published
    April 13, 2021, 1:15 p.m.
    ...strain. In our metabolic engineering case study of N-acetylglucosamine biosynthesis, we constructed a strain...) with 4 rounds of transformation to complete the N-acetylglucosamine strain library construction in 8...cargo copy numbers. (D) The effect of gene dosage on N-acetylglucosamine concentration. Image from the Yang... clearance. It is more convenient to update the spacer(20nt) on pEcgRNA plasmid as only two 24 nt oligos... proteins Cas6, Cas7, Cas8, and the crRNA. The spacers of the crRNA can be customized as well, including...

  31. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

    Type
    Blog Post
    Published
    Sept. 3, 2019, 1:15 p.m.
    ...deposit please sign up here.        New toolkit for N or C terminal tagging of proteins for degradation ...lab has created a new toolkit of plasmids for the N- or C- terminal tagging of proteins with mAID and ...can be limited by the need for a specific PAM (protospacer adjacent motif) in order to bind the target sites...These loops not only provide a 3-D structure for packing genomic DNA into nuclei, but are also thought to...

  32. Components of CRISPR/Cas9

    Type
    Blog Post
    Published
    Feb. 2, 2016, 5 p.m.
    ...and its variants have two endonuclease domains: the n-terminal RuvC-like nuclease domain and the HNH-like...Staphylococcus aureus (SA); SaCas9 3' NNGRRT or NNGRR(N)  Acidaminococcus sp. (AsCpf1) and Lachnospiraceae...RNA provided that target is upstream (5') of a protospacer adjacent motif (PAM) Cas9 endonuclease binding... RNA and a 3-base pair sequence known as the Protospacer Adjacent Motif or PAM. In order for dsDNA to ... and the unique stretches of DNA between them “spacers” (see figure below). After years of research, we... each direct repeat, combined with its adjacent spacer, ultimately encodes a single crRNA. The direct ...pre-crRNA into mature crRNA and tracrRNA binding. The spacer regions, on the other hand, are the unique, foreign...

  33. CASTing Off for New Shores in Human Genome Editing

    Type
    Blog Post
    Published
    June 10, 2025, 1:15 p.m.
    ...Eitzinger, S., Miller, S. M., Berríos, K. N., McElroy, A. N., King, R. T., Stringham, O. G., Gelsinger...power of CRISPR combined with the large sequence capacity of transposons open whole new worlds to explore...

  34. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    Published
    May 5, 2020, 1:15 p.m.
    ...2020.02.29.971127 Rauch, J. N., Valois, E., Solley, S. C., Braig, F., Lach, R. S., Baxter, N. J., Kosik, K. S.,...., Phutela, R., Ansari, A. H., Sinha, D., Sharma, N., Kumar, M., … Maiti, S. (2020). Rapid, field-deployable...the target site, while Cas12a requires a PAM (Protospacer Adjacent Motif) for target cleavage. This test...

  35. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ...-07259-6 Yarnall, M. T. N., Ioannidi, E. I., Schmitt-Ulms, C., Krajeski, R. N., Lim, J., Villiger, L.,...s41587-021-01039-7 Pandey, S., Gao, X. D., Krasnow, N. A., McElroy, A., Tao, Y. A., Duby, J. E., Steinbeck...Zhou, W., Jiang, K., Garushyants, S. K., Roberts, N., Zhang, L., Vakulskas, C. A., Walker, J. A., Kadina... formation slightly. Designing the sgRNA with a spacer that only binds the edited strand, as in the PE3b...Phage-assisted evolution and protein engineering yield compact, efficient prime editors. Cell, 186(18), 3983-4002...

  36. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    Published
    June 4, 2024, 1:15 p.m.
    ...Cas9 recognizes the PAM sequence 5’-NGG-3’, where N can be any base. In Figure 5, we can see why: two ...-specific hydrogen bonds to the guanines, but the N (a thymine in this case) is only bound by the phosphate...References Bravo, J. P. K., Liu, M.-S., Hibshman, G. N., Dangerfield, T. L., Jung, K., McCool, R. S., Johnson...Hsu, P. D., Konermann, S., Shehata, S. I., Dohmae, N., Ishitani, R., Zhang, F., & Nureki, O. (2014). Crystal...important to choose a target sequence with a nearby protospacer-adjacent motif (PAM)? Cas9 is becoming an ever...

  37. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    Published
    Feb. 24, 2015, 7:50 p.m.
    ..., Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O. Cell. 2014 Feb 27;..., Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F. Nature...gain-of-function screening when combined with a compact pooled sgRNA library. Our SAM-mediated screens ...

  38. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    Published
    Jan. 5, 2021, 2:15 p.m.
    ... light, Vvd dimerizes bringing the C-terminal and N-terminal Cre fragments together and permitting recombinase...reduced RNA-off target footprint compared to current N-terminal linked SaCas9 ABE variants. It's also one.... https://pubmed.ncbi.nlm.nih.gov/24390935/ Hypercompact genome editor CRISPR-Casɸ By Melina Fan The ...toolbox with their discovery of CRISPR-Casɸ, a hypercompact genome editor from the Biggiephage clade of ...the Doudna Lab. Scientists looking for more compact genome editors due to delivery constraints and ...

  39. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    Published
    Aug. 29, 2024, 1:15 p.m.
    ... promoters, N’ and/or C’ terminus, and selection markers. Need fewer components? A spacer can be inserted...L., Szegletes, Z. M., Wen, B., Kraus, C., Miller, N. W., Steger, R. J., Escude Velasco, B., Bosch, J. ...entirely, but instead will have to replace it with a spacer.) Each component available on the Fragmid website...

  40. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    Published
    March 21, 2024, 1:15 p.m.
    ...resources   References Allikalt, A., Purkayastha, N., Flad, K., Schmidt, M. F., Tabor, A., Gmeiner, P....Patel, R., Lu, R., Macklin, J. J., Keller, P. J., Ji, N., & Lavis, L. D. (2017). A general method to fine-...Hayashi-Takanaka, Y., Stasevich, T. J., Kurumizaka, H., Nozaki, N., & Kimura, H. (2014). Evaluation of chemical fluorescent...realizing it. Yet, this selection is crucial as it will impact downstream outcomes of an experiment, including...Grimm & Lavis, 2022). These variations directly impact the way you can utilize these dyes. The most-used...emission of labeled agonists and antagonists, impacting the interpretation of experimental results. It's...dopamine D2/D3 receptors (Allikalt et al., 2020). Impact of labeling process: The process of dye labeling...
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