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We narrowed to 114 results for: N-pac

Showing: 81 - 114 of 114 results

  1. Working with Nuclear Receptors

    Type
    Blog Post
    Published
    July 15, 2014, 4:03 p.m.
    ...regulation. Molecular Cell 27, 691-700. (PubMed) McKenna, N. J.and O'Malley, B. W. (2002). Combinatorial control...cascades conceivably allows the genome to sense the impact of the total environment on the cell. The NURSA...

  2. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    Published
    Aug. 7, 2018, 12:31 p.m.
    ...Kumar M, Keller B, Makalou N, Sutton RE. Systematic determination of the packaging limit of lentiviral vectors...generation packaging systems use 3 plasmids; an envelope plasmid that usually encodes VSV-G, a packaging plasmid...lentiviral packaging systems please see Addgene’s Lentiviral Guide. While the third generation packaging systems...requirement for additional DNA can impact the transfection efficiency of the packaging cell line and thereby affect...are transfected into a packaging cell line, usually a derivative of 293T. Packaging cells are incubated ...delivering genes via lentivirus is deciding what packaging system is best for a particular experiment. The...The newer generation lentiviral packaging systems were designed to prevent the formation of replication...

  3. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    Published
    April 14, 2022, 1:15 p.m.
    ...Chemistry. https://doi.org/10.1074/jbc.M407888200 Kwon, N., Kim, Y., Lee, J. (2019). Structural diversity and...amazing affinity reagents we just didn't have the space to give proper credit to. Thankfully, as Addgene's..., or be able to access an epitope folded into a space that would size-exclude an antibody. In imaging ...

  4. Antibodies 101: Fab Fragments

    Type
    Blog Post
    Published
    Sept. 5, 2024, 1:15 p.m.
    ...Sakai M, Tomisaki M, Kiuchi T, Takagi J, Watanabe N (2022) Engineered fast-dissociating antibody fragments...sometimes you want a tool that is a little more compact — more of an antibody fragment rather than the ...

  5. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    Published
    April 28, 2016, 2:30 p.m.
    ...information. References: 1. Schnütgen F, Doerflinger N, Calléja C, Wendling O, Chambon P, and Ghyselinck ...and Dymecki, SM. Talking about a Revolution: The Impact of Site-Specific Recombinases on Genetic Analyses...

  6. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...Vidigal JA, Han Y-C, Ogrodowski P, Crippa A, Rekhtman N, de Stanchina E, Lowe SW, Ventura A (2014) In vivo...provide a large transgene packaging capacity. For instance, with a packaging capacity of about 6 kb, the 1st...of AdVs have already a packaging capacity larger than AAV vectors. This capacity is large enough to carry...

  7. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    Published
    May 7, 2020, 1:15 p.m.
    ...to contain a specific protospacer adjacent motif (PAM) made up of 5′-NGG-3′ (“N” can be any nucleotide... Depending on the position in the protospacer, the editing capacity of the ABE8s ranged from 1.5x higher...ABE8s demonstrate an overall improved base editing capacity, even at sites that used to be difficult to target....  References Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson DI, Liu DR (2017) Programmable...Edwards A, Gehrke JM, Lee S-J, Liquori AJ, Murray R, Packer MS, Rinaldi C, Slaymaker IM, Yen J, Young LE, Ciaramella...

  8. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    Published
    June 12, 2014, 3:30 p.m.
    ...plants in mind, Arabidopsis and Nicotiana benthamiana. N. benthamiana is a tobacco relative and a good model...other people to do that too. We don’t have the capacity to send tools out to people ourselves, but we’...committees, and how the culture of the university impacts the number of women reaching senior positions. ... those amazing students that want to have a big impact on the world and tell them to work in plant science...

  9. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...21439297. PubMed Central PMCID: PMC3622210. Schek N, Cooke C, Alwine JC. Definition of the upstream efficiency...(A) signals in combination with certain viral packaging systems has been associated with reduced viral...

  10. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ...References 1. Aurnhammer, C., Haase, M., Muether, N., Hausl, M.A., Rauschhuber, C.T., Huber, I., Nitschko...analyses needed for packaging AAV. AAV production Overview: The first step in packaging AAV is transfecting...had never worked with AAV, so naturally I had to package some virus for my first experiment. It was a bit...transfecting HEK293 cells with AAV packaging plasmids. The cells need three different plasmids to produce AAV...intermediate into the single-stranded genome that's then packaged into AAV’s protein shell, or capsid; 2) the pHelper...plasmid which encodes a transgene of interest that’s packaged into the virus. Two to five days after transfection...plasmids contain ITRs which are required for proper packaging of AAV, but also form secondary structures that...

  11. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    Published
    May 15, 2014, 3:33 p.m.
    ... of the most common uses, GFP can be fused to the N- or C-terminus of a protein, which allows the scientist...from quenching by aqueous solvent. This tightly-packed structure explains the importance of the entire...stability, GFP can be used in lineage tracking capacities in cell fate studies. It can also be used, when...

  12. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    Published
    Aug. 9, 2016, 2:30 p.m.
    ...GENOME --local -f -k 10 --very-sensitive-local -L 9 -N 1 -U GRNA_23MERS -S GRNA_HITS.sam Many thanks to ...action at a distance.” Through the marvel of DNA compaction, an enhancer can increase the expression of a...millions of base pairs away. (For details on DNA compaction and the structural proteins that make it possible...details. DNA is a 3D Fractal Figure 2: DNA compaction. Graphics courtesy of beyondthedish (left) and...tertiary structures (globules) to achieve exquisite compaction—on the order of 700 terabytes per gram—and form...chromosomes we all know and love. To understand compaction, suppose you have a piece string that’s 10 meters..., are mere millimeters apart. Similarly, cells compact DNA in a way that brings linearly distant regions...

  13. Bioinformatics at Addgene

    Type
    Blog Post
    Published
    July 14, 2022, 1:15 p.m.
    ... filtered the input channel to only use the first n items. To be sure, we went through much trial and ...complete package, all of the pieces of software that depend on each other for the entire package to run....in-house software has been crucial for us to keep pace with our current need to analyze nearly 300 sequences...sequencing data collection and analysis eventually outpaced our software, even with continuous updates. The...

  14. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    Published
    Aug. 27, 2020, 1:15 p.m.
    ...information, you can fuse each FP fragment to the N- and C-terminal end of each PIP and empirically test...sequences, the amino acid sequences RSIAT and RPACKIPNDLKQKVMNH have been used successfully (Kerppola, 2006...expressed transiently by transfection or stably impacts the success of BiFC experiments. In either case...//doi.org/10.1101/pdb.prot076497 Kim Y, Jung JP, Pack C-G, Huh W-K (2018) Global analysis of protein homomerization...

  15. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    Published
    Dec. 7, 2020, 2:53 p.m.
    ...M, Zhou Z, Smock DC, Tang K, Owens TK, Krishnappa N, Sachdeva R, Deutschbauer AM, Banfield JF, Doudna ...top):  A CRISPR RNA (crRNA) comprising a 32 nt spacer sequence specifying the target site. Four proteins...INTEGRATE using a single plasmid containing a multi-spacer CRISPR array. Workflow and technical considerations...INTEGRATE system follows a simple workflow: Clone spacer(s) into the CRISPR array Clone desired DNA cargo...

  16. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    Published
    Nov. 14, 2024, 2:15 p.m.
    .... 2015;6. doi:10.3389/fpls.2015.01001 Ho TT, Zhou N, Huang J, et al. Targeting non-coding RNAs with the...start site function would also help to reveal the impacts of a lncRNA on nearby genes and its role in gene...

  17. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    Published
    March 28, 2018, 6:52 p.m.
    ...genomes. Cas9s from other species, like S. aureus and N. meningitidis require different PAM sites than SpCas9... 64 possible NNN PAM sequences adjacent to a protospacer. If ω-dCas9 could recognize a library PAM, it...

  18. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Published
    July 31, 2018, 12:55 p.m.
    ...Central PMCID: PMC4267879. 10. Samulski RJ, Muzyczka N., 2014. AAV-Mediated Gene Therapy for Research and...integration into the genome, and its large DNA packaging capacity is well suited for larger transgenes. Unfortunately...

  20. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    Published
    Nov. 16, 2021, 2:15 p.m.
    ...https://doi.org/10.1007/978-1-4939-8935-5_26 Stuurman N. (2012) Introduction to Fluorescence Microscopy. iBiology...architecture of my cells,” or “How does this mutation impact the types of cells found in my tissue.” It is based...antibody performs differently and some antibodies impact the performance of other antibodies. But, once ...the right protocol, multiplex IF can give you impactful and beautiful results. Performing an immunofluorescence...fixation maintains sample morphology with minimal impact to the target epitope. There are multiple methods...exclusive to IF, but they can have a significant impact on the outcome of your IF experiment. Many of the...choose for your experiment will have a significant impact on the protocol you use and the ultimate outcome...

  21. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    Published
    Oct. 13, 2022, 1:45 p.m.
    ...2017).Walton, R. T., Christie, K. A., Whittaker, M. N. & Kleinstiver, B. P. Unconstrained genome targeting... target is present immediately upstream of a Protospacer Adjacent Motif (PAM), the sequence NGG.  If these... SpCas9 and its enhanced variants, eSpCas9 and HypaCas9, are great choices. Check out our SpCas9 containing...Cas9 variant with higher specificity (eSpCas9 and HypaCas9) instead.  Choosing a Cas9 for site-specific modifications... within the editing window (bases 4-8 of the protospacer). While this is a small window, this system can...

  22. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    Published
    April 7, 2016, 2:30 p.m.
    ...(2009): 495-505. PubMed PMID: 19300443. 7. Holic, N., Seye, A.K., Majdoul, S., Martin, S., Merten, O.W...lentiviral infection begins with your cells, both your packaging cells and your target cells. Poor tissue culture...harvest after transfecting with the transfer and packaging plasmids. A benefit of the multiple harvest strategy...benefitting your cells, medium exchanges may also impact the quality of the lentiviral particles you harvest...density, volume, incubation time, and MOI can all impact the success of your lentiviral infection. To ensure...

  23. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    Published
    Nov. 19, 2020, 2:15 p.m.
    ...Dennys-Rivers C, Huffenberger A, Braun L, Rinaldi F, Wein N, Meyer KC, Solano S, Nguyen K , Lang E, Kaspar AA,...trans-splicing, and protein replacement to reverse the impacts of this disease.  As a non-degenerative, monogenic...

  24. Antibodies 101: Normalization and Loading Controls for Western Blots

    Type
    Blog Post
    Published
    April 8, 2025, 1:15 p.m.
    ...membrane versus the amount of protein loaded on the gel (n=3). Data is presented as mean ± SEM. Adapted from ...controls only address one type of error that can impact your western blot, and depending too much on normalization...

  25. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    Published
    Feb. 8, 2024, 2:15 p.m.
    ...likelihood of an off-target artifact. Increasing the N in your experiment is the only way to effectively ...in mind, but likely will not have a significant impact on your data. You are trying to generate a knockout...depending on your platform, off-target effects should impact your experimental design differently. Yes, it is...optimization which have led to more specific variants. HypaCas9, sSpCas9(1.1), SpCas9HF1, and evoCas9 are a few...

  26. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    Published
    Sept. 29, 2020, 1:15 p.m.
    ... https://doi.org/10.1242/dmm.002733 Idalia, V.-M. N., & Bernardo, F. (2017). Escherichia coli as a Model...more model organisms that have left a historical impact on the scientific community. (And, check out this...

  27. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    Published
    June 24, 2020, 5:45 p.m.
    ...10.1038/onc.2014.470 Taghbalout A, Du M, Jillette N, et al (2019) Enhanced CRISPR-based DNA demethylation...modifications to DNA or histones, the proteins that help package DNA, were associated with gene expression or silencing...particularly large protein, and this fusion allows for packaging into viral vectors and targeted delivery into ...

  28. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Sept. 24, 2020, 1:15 p.m.
    ...avoid target sites that code for amino acids near the N’ terminus of the protein, in order to mitigate the...thus frameshifts that disrupt the protein-coding capacity of a locus. When using S. pyogenes Cas9 (SpCas9.../doi.org/10.1038/s41587-020-0561-9 Dempster JM, Pacini C, Pantel S, Behan FM, Green T, Krill-Burger J,...

  29. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    Published
    July 28, 2016, 2:30 p.m.
    ...biocomputing and biological circuits.     References 1. N. Roquet et al., Synthetic recombinase-based state ...are composed. Fear not – the state machine design space is highly degenerate and often times there are multiple... et al. "Permanent genetic memory with> 1-byte capacity." Nature methods 11.12 (2014): 1261-1266. PubMed...

  30. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    Published
    Nov. 2, 2021, 1:15 p.m.
    ...Williams P, Zolotukhin S, Reier PJ, Mandel RJ, Muzyczka N (2004) Recombinant AAV Viral Vectors Pseudotyped with...recombination between homologous pairs before viral packaging even begins. This spontaneous recombination persists...2020). In general, we observe that 0.1 - 0.8% of packaged viral genomes contain a recombined/functional ...which can lead to off-target expression of the packaged transgene in neurons and glia.   Check the recombination...v14715, titer 2.4x10e13 GC/mL) for this study were packaged at Addgene and had a relatively low recombination...

  31. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ...transposons-the-jumping-genes-518/   Sandoval-Villegas N, Nurieva W, Amberger M, Ivics Z (2021) Contemporary...between the transposon’s LTRs, similar to when packaging viral vectors.   Let’s take a closer look at three...after cleavage by the transposase. It has a cargo capacity >100 kB, although integration efficiency decreases...without decreased efficiency, with a maximum cargo capacity of ~200 kB of DNA. Similar to piggyBac, Tol2 also...

  32. Antibodies 101: Validation

    Type
    Blog Post
    Published
    March 24, 2022, 1:15 p.m.
    ... Lisnic VJ, Kremer L, Li D, Koch-Nolte F, Pascual N, Rodríguez-Barbosa J-I, Torensma R, Turley H, Pulford...specificity. However, there are countless variables that impact an antibody’s affinity and specificity for a given...

  33. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...The pCoofy series of plasmids contain a variety of N- and C-terminal tags (including His, S-tag, OneStrep...types of pre-cloned insert modules (plant promoter, N-terminal tag, coding sequence of the gene of interest...optimal scaffold can depend on the length of the spacer region between the target sequences. These updates...

  34. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ...drop-down menu to indicate whether the tag is on the N or C terminus and whether it was present in the backbone...with a unique Deposit ID. Addgene will send you a package containing instructions on how to prepare your ...
Showing: 81 - 114 of 114 results