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We narrowed to 114 results for: N-pac

Showing: 41 - 80 of 114 results

  1. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    Published
    May 19, 2016, 2:30 p.m.
    ... J, Larghero J, Kabbara N, Dalle B, Gourmel B, Socie G, Chrétien S, Cartier N, Aubourg P, Fischer A, Cornetta...of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid carried a mutated...Advantages of lentivirus include a large genetic capacity and the ability to transduce both dividing and...mutated HIV-1 provirus that couldn’t package itself due to a few missing proteins. The envelope plasmid ...terminal repeats (LTRs) that facilitate viral packaging and host genome integration. After co-transfection...the brain (Figure 1). 2nd generation lentiviral packaging system The graphic below shows how the lentiviral... infect CD4+ cells. 3rd generation lentiviral packaging system Although the 2nd generation system is safer...

  2. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    Published
    Feb. 4, 2025, 2:15 p.m.
    ...Muller, A., Sousa, A. A., Tsai, S. Q., Bengtsson, N. E., Lööv, C., Ingelsson, M., Chamberlain, J. S., ...j.ymthe.2022.09.015 Ramani, B., Rose, I. V., Teyssier, N., Pan, A., Danner-Bocks, S., Sanghal, T., Yadanar,...to using AAV is the limited packaging capacity. Lentiviruses have capacities of 8–12 kb, while AAV only...libraries available at Addgene. The large packaging capacity and stable expression of lentiviral vectors...Lentivirus has limited packaging options, with most lentiviral vectors being packaged using VSV-G. In contrast...contrast, AAV can be packaged in a variety of capsids. To date, at least 12 different capsids, or serotypes...that when using Cas9, you’re already almost out of space! Don’t let this get you down for too long, though...

  3. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...annurev-biophys-083012-130404 Zhang N, Zhang D, Chen SL, Gong B-Q, Guo Y, Xu L, Zhang X-N, Li J-F (2018) Engineering...tpc.112.098509 Smits AH, Ziebell F, Joberty G, Zinn N, Mueller WF, Clauder-Münster S, Eberhard D, Fälth ...the most prevalent method due to ease of use and capacity to generate a long-term RNAi phenotype. Long hairpin...

  4. Common Injection Routes in Mice

    Type
    Blog Post
    Published
    Jan. 22, 2025, 9:42 p.m.
    ...liquid. Restraint: anesthesia Location: nose Gauge: n/a Volume: < 0.05 ml Skill required: beginner Oral ...) < 0.07 mL Advanced Intranasal Anesthesia Nose n/a < 0.05 mL Beginner Oral gavage Scruffing Throat...injections are given in the peritoneal cavity, or the space surrounding organs in the abdomen. They are a popular...cerebral spinal fluid. The injection site is the space in and around the vertebrae, so hitting the correct...

  5. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...PubMed PMID: 4355596 King, S. R., Duggal, N. K., Ndongmo, C. B., Pacut, C., & Telesnitsky, A. (2008). Pseudodiploid...Teich, N. (1971). Noninfectious ARK mouse embryo cell lines in which each cell has the capacity to be ...the genetic code for reverse transcriptase are packaged (Wu et al., 1997) inside of the capsid . This ...the vector particle, some of the RT protein gets packaged inside of the vector particle, along with its ...-type viral genome can direct the production, packaging, and release of progeny viral particles. And there... Temin, H. M. (1990). Genetic consequences of packaging two RNA genomes in one retroviral particle: pseudodiploidy...

  6. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    Published
    Feb. 27, 2018, 2:04 p.m.
    ...Antibody Subclass Nanobody Addgene Plasmid Rabbit N/A* TP897 pTP1183 (1x cysteine) pTP955 (3x cysteines.... Nanobodies will bind to the column due to their N-terminal His14-bdNEDD8 tag. After several washes, ...antibody (HCab), which are unique to camels, llamas, alpacas and other camelids. In terms of structure, HCabs...have done the hard work for you by immunizing an alpaca, isolating HCab antibodies from its serum, and ...for generating nanobodies are outlined below; no alpaca required. Decide Which Nanobody To Use: The table...

  7. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    Published
    Feb. 17, 2022, 2 p.m.
    ...Eleonora I. Ioannidi, Matthew T. N. Yarnall, Cian Schmitt-Ulms, Rohan N. Krajeski, Justin Lim, Lukas Villiger...discovery, including CRISPR, is turning potential into impact. In molecular and cellular biology, this happens...Jonathan Gootenberg, PhD, focuses exclusively on this space — developing tools for cellular targeting. Lately...and cleaving RNA at positions defined by the target:spacer duplex without detectable non-specific activity...

  8. Using AAV for Neuronal Tracing

    Type
    Blog Post
    Published
    Aug. 9, 2018, 1:04 p.m.
    ...Saunders, A., Oldenburg, I.A., Miyamichi, K., Akhtar, N., Luo, L., Whelan, S.P., Sabatini, B., and Cepko, ...K.Y., Jang, M.J., Yoo, B.B., Greenbaum, A., Ravi, N., Wu, W.-L., Sánchez-Guardado, L., Lois, C., Mazmanian...Kitamura, T., Suh, J., Frank, D., Kajikawa, K., Mise, N., Obata, Y., Wickersham, I.R., and Tonegawa S. (2014...Oh, S.W., Harris, J.A., Ng, L., Winslow, B., Cain, N., Mihalas, S., Wang, Q., Lau, C., Kuan, L., Henry,... applied, but these methods can have a negative impact on cellular health. Another limitation of traditional... them capable of diffusing in the extracellular space; they also have low immunogenicity and cellular ...less likely to be sequestered in the extracellular space. In addition, rAAV2-retro’s altered amino acid sequence...

  9. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    Published
    June 25, 2024, 1:15 p.m.
    ...https://doi.org/10.1016/j.cell.2024.03.020 Mathis, N., Allam, A., Kissling, L., Marquart, K. F., Schmidheini...https://doi.org/10.1038/s41587-022-01613-7 Mathis, N., Allam, A., Tálas, A., Kissling, L., Benvenuto, E.... The pegRNA consists of a target-complementary spacer, a scaffold sequence, and an extension sequence.... L., Medema, R. H., & van Steensel, B. (2021). Impact of chromatin context on Cas9-induced DNA double-strand...

  10. Hot Plasmids Spring 2024

    Type
    Blog Post
    Published
    May 23, 2024, 1:15 p.m.
    ...developed a new prime editor that incorporates La’s N-terminal domain, PE7 (Figure 2).  PE7 shows enhanced...-CoV-2 nucleocapsid protein (Plasmid #141391-LV [+N]) or membrane protein (Plasmid #141386-LV [+M], negative...single-strand AAV (ssAAV). To maintain the packaging capacity and improve the specificity of ssAAV, a targeting...CRISPRi screen to identify additional host factors impacting prime editing and found the protein La (a small...proteins and a critical component for viral genome packaging. Addgene recommends both for use in western blots...that was more desirable than having many closely-packed labeled cells. Plus, Voltron2’s fluorescence is...vectors for microglia was further enhanced by packaging the virus as self-complementary AAV (scAAV). However...

  11. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

    Type
    Blog Post
    Published
    July 12, 2021, 1:15 p.m.
    ...Johnson NV, Ludes-Meyers J, Nguyen AW, Park J, Wang N, Amengor D, Lavinder JJ, Ippolito GC, Maynard JA, ...P, Gautam A, Windsor I, Travers M, Chen Y, Garcia N, Whiteman MB, McKay LGA, Lelis FJN, Habibi S, Cai ... are cloned into pLVX vectors (for lentiviral packaging) and contain an IRES-Puro marker for stable cell...

  12. Hot Plasmids: Winter 2025

    Type
    Blog Post
    Published
    March 11, 2025, 1:15 p.m.
    ... prime editing plasmids here! Vu, T. V., Nguyen, N. T., Kim, J., Song, Y. J., Nguyen, T. H., & Kim, J...double-tagged constructs (including "CLAP", combining an N-terminal CLIPf tag and a C-terminal SNAPf tag; see...from a vector backbone that allows lentiviral packaging as a single payload (Figure 4). Figure...

  13. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    Published
    Aug. 20, 2020, 2:30 p.m.
    ...Find plasmids from this paper at Addgene. Amrani N, Gao XD, Liu P, Edraki A, Mir A, Ibraheim R, Gupta... Hu JH, Miller SM, Geurts MH, Tang W, Chen L, Sun N, Zeina CM, Gao X, Rees HA, Lin Z, Liu DR (2018) Evolved...) the sequence is located just upstream of a Protospacer Adjacent Motif (PAM sequence). The 3-5 nucleotide...about 1 kilobase smaller than SpCas9, so it can be packaged into adeno-associated virus (AAV).  At 984 amino...edit. Some Cas13 enzymes require a single base protospacer flanking sequence (PFS) adjacent to the target...

  14. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    Published
    July 7, 2020, 1:15 p.m.
    ...9 St. Martin A, Salamango D, Serebrenik A, Shaban N, Brown WL, Donati F, Munagala U, Conticello SG, Harris...nar/gky332 Standage-Beier K, Tekel SJ, Brookhouser N, Schwarz G, Nguyen T, Wang X, Brafman DA (2019) A ... The lab then introduced an sgRNA targeting the GFPACG to the cells. As expected, only the cells that ...luciferase. References Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson DI, Liu DR (2017) Programmable...

  15. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    Published
    May 13, 2025, 1:15 p.m.
    ...PMC2496964 Leavitt, A. D., Robles, G., Alesandro, N., & Varmus, H. E. (1996). Human immunodeficiency virus... Central PMCID: PMC449739 Yew, C.T., Gurumoorthy, N., Nordin, F., Tye, G.J., Wan Kamarul Zaman, W.S., ...further! You can package your own lentiviral vector using an integrase-deficient packaging plasmid available... the integrase-deficient packaging plasmid instead of the standard packaging plasmid. For more details...integration of this portion into the host genome. The packaging plasmid contains Gag, Pol, Rev, and Tat genes....transcriptase), and IN (integrase). These viral packaging genes are required for the production of lentiviral...integrated into the host genome via the viral proteins packaged during production (read our blog post on the retroviral...

  16. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    Published
    Nov. 3, 2022, 12:15 p.m.
    .../doi.org/10.1093/nar/gku936 Eva K. Brinkman, Arne N. Kousholt, Tim Harmsen, Christ Leemans, Tao Chen, ...10.1093/nar/gky164 Cancellieri S, Canver MC, Bombieri N, Giugno R, Pinello L. CRISPRitz: rapid, high-throughput... editing, such as SpCas9-HF1, eSpCas9(1.1), or HypaCas9. Read more about Cas9 options in our blog post...

  17. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    Published
    June 17, 2025, 1:15 p.m.
    ..., P., Ashoor, H., Åström, G., Babina, M., Bertin, N., Burroughs, A. M., Carlisle, A. J., Daub, C. O., ...Han, J., Yeom, K., Lee, S., Baek, S. H., & Kim, V. N. (2004). MicroRNA genes are transcribed by RNA polymerase...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...

  18. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    Published
    April 24, 2015, 2:39 p.m.
    ...Brainbow-2.1 construct can express one of four colors (n-GFP, RFP, YFP or M-CFP.) The construct contains two...van Es JH, van den Born M, Kroon-Veenboer C, Barker N, Klein AM, van Rheenen J, Simons BD, Clevers H. Cell...development of a new technique. In a structure as tightly packed and interwoven as the brain, robust single-cell...

  19. Easing the Protein Purification Process with pCri

    Type
    Blog Post
    Published
    June 19, 2015, 3:08 p.m.
    ...includes plasmids with different varieties of tags, both N- and C-terminal. Each set of plasmids uses the same...yeast Pichia pastoris. The improved secretory capacities of these microbes make them useful for producing...difficult-to-fold proteins. In addition, P. pastoris has the capacity to glycosylate proteins and may recapitulate the...

  20. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    Published
    May 31, 2016, 3:30 p.m.
    ...27075976. 4.  X. Li, E. R. Burnight, A. L. Cooney, N. Malani, T. Brady, J. D. Sander, J. Staber, S. J. ...Joung, P. B. McCray, F. D. Bushman, P. L. Sinn, and N. L. Craig, “piggyBac transposase tools for genome ...Furthermore, piggyBac™ offers a large cargo-carrying capacity (over 200 kb has been demonstrated) with no known...certain genes, such as those that have negative impacts on fitness. One consideration when making a transgenic...

  21. Reflections and Looking Ahead

    Type
    Blog Post
    Published
    Feb. 25, 2025, 2:15 p.m.
    ...Virk, H., Laflamme, C., Houston, D. W., Polinski, N. K., Meijers, R., Levey, A. I., Saper, C. B., Errington...these milestones remind us of the extraordinary impact that happens when scientists come together to share...more AAV viral vector options, we launched AAV Packaged on Request, a service that dramatically expands... transfer plasmid in our repository can now be packaged on demand in five distinct serotypes, offering...breakthrough enabled brings us closer to making an impact on humanity's greatest challenges. Thank you for...

  22. Antibodies 101: Fc Effector Functions

    Type
    Blog Post
    Published
    May 18, 2023, 1:15 p.m.
    ...10.1038/cr.2009.139 Janda, A., Bowen, A., Greenspan, N. S., & Casadevall, A. (2016). Ig Constant Region Effects...bind. We also know that changing the Fc domain can impact an antibody's binding affinity (Janda et al., 2016...

  23. An Addgene Summer Internship

    Type
    Blog Post
    Published
    Aug. 8, 2024, 7:22 p.m.
    ... Syntaxin3, and osa-miR528. My manager, Katherine N., reviewed the list and followed up with her manager...first week at Addgene, I had to get used to the workspaces, as well as complete the lab safety and cybersecurity...safety, general work skills — such as using Google workspace — and got the chance to work on my very own research...

  24. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    Published
    Nov. 25, 2013, 10:12 p.m.
    ... Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini LA, Zhang F. Science...blockbuster known as the CRISPR (clustered regularly interspaced palindromic repeats) Cas (CRISPR associated genes...genetic material next to the species-specific protospacer adjacent motif (PAM). Once the CRISPR/Cas complex...

  25. Another Pathway into Cells: iTOP

    Type
    Blog Post
    Published
    June 23, 2015, 8:37 p.m.
    ...Clevers H, Prasad V, Lebbink RJ, Rehmann H, Geijsen N. Cell. 2015 Apr 23;161(3):674-690. doi:10.1016/j.cell...to a protein of interest thereby giving it the capacity to enter cells. One such CPP was discovered in...directly with proteins removes the limitations of packaging size associated with viral vectors. For these ...

  26. Revamp Your Lab Meetings With Creative Virtual Collaboration

    Type
    Blog Post
    Published
    Feb. 9, 2021, 2:15 p.m.
    ..., Weyhenmeyer GA, O’Reilly CM, Lofton ME, Catalán N (2018) Ten simple rules for collaboratively writing...and novelty in scientific research and in any workspace. Re-design lab meetings with a focus on discussion...making lab meetings more interactive and creating space for discussion at the end of each talk - time for...

  27. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    Published
    Sept. 21, 2021, 1:15 p.m.
    ... four different categories.     Brookhouser, N., et al. BMC Biol 18, 193 (2020). https://doi.org/...several pooled libraries: Cas13X and Cas13Y, two compact Cas13 proteins found in the metagenome datasets...

  28. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    Published
    June 2, 2015, 6:06 p.m.
    ...highly specific genome editing. Tsai SQ, Wyvekens N, Khayter C, Foden JA, Thapar V, Reyon D, Goodwin MJ...Cas9 multiplexing in systems with limited packaging capacity. Still confused about the various tRNA expression... will be necessary to improve its multiplexing capacity. Multiplexing is key due to the natural redundancy...often necessary. Guide RNAs (gRNAs) are commonly packaged in 400-500 bp cassettes containing the RNA pol...100 bases) limit the number of gRNAs that can be packaged together in a single vector. In addition, the ...forms tRNA-gRNA and tRNA-gRNA-tRNA, with short 5’ spacer sequences preceding gRNAs, to test for gRNA cleavage...predicted with no addition of nucleotides to the 5’ spacers. The 3’ ends were modified slightly with either...

  29. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    Published
    April 16, 2015, 3:45 p.m.
    ... Center at UNC Chapel Hill.   References Bennardo N, Gunn A, Cheng A, Hasty P, Stark JM (2009) Limiting...experiment. Here, we’ll discuss NHEJ, and how it impacts what happens to Cas9-mediated DSBs in the genome...active throughout the cell cycle and has a higher capacity for repair, as there is no requirement for a repair...chromosomes within a day. Another factor expected to impact repair is that the Cas9 protein doesn’t immediately...in determining how the structure of broken ends impacts how they are repaired. He works in Dale’s lab.  ...

  30. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ....The actual oligo you should order is:5'-CACCGN...N-3'3'-CN...NCAAA-5'You need to add the CACC and AAAC...Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P. D., Wu, X., Jiang, W., Marraffini, L. A...."CRISPR", short-hand for Clustered Regularly Interspaced Short Palindromic Repeats, in Pubmed returned...want to use double nickase, you could express two spacers and use PX335 to express the Cas9n (nickase). The...PAM sequence to the oligo when cloning my target spacers into the PX330 vector backbone? A2: There is no...usually adds it when they order the oligo. If your spacer sequence starts with a 'G', you naturally have ... compared to where the target site sequence (protospacer+PAM) is located? If there are other PAMs in the...

  31. How Dry Ice Affects Viral Vector Sample pH & How to Avoid It

    Type
    Blog Post
    Published
    June 20, 2017, 2:30 p.m.
    ...Chipman P, Agbandje-McKenna M, Cleaver BD, Clément N, Byrne BJ, Zolotukhin S. A simplified purification...keep viruses frozen during shipment was having an impact on the samples. We therefore devised a few experiments...solution thawed, as the CO2 was confined to the tube headspace when the samples were frozen. This was clear by...tubes prior to thaw released the CO2 from the headspace, thus mitigating the pH effects of dry ice storage...

  33. Viral Vectors 101: Biosensors

    Type
    Blog Post
    Published
    July 6, 2023, 1 p.m.
    ... you covered with packaging plasmids! References and Resources References  Bhalla N., Jolly P., et al...with some of our most popular biosensors already packaged and ready to deliver. Ready to give biosensors...

  35. Summer SciComm: Preprints

    Type
    Blog Post
    Published
    Aug. 16, 2022, 1:15 p.m.
    ... Open. https://doi.org/10.1242/bio.059310 Fraser N, Mayr P, Peters I (2021) Motivations, concerns and...scientists know the results, your work won’t have any impact! There are lots of ways scientists communicate...efforts, inspire future research, and increase their impact. Open access and transparency Another major benefit...

  36. Deep Dive: qPCR

    Type
    Blog Post
    Published
    Sept. 29, 2022, 1:15 p.m.
    ....2008.112797. Epub 2009 Feb 26. PMID: 19246619.Downey N. Interpreting melt curves: An indicator, not a diagnosis...reaction set-up. Nucleic Acids Res. 31:e122.Thelwell N; Millington S; Solinas A; Booth J; Brown T. Mode of...multiplex your reference genes to save on plate space. For many cell lines and tissue types, appropriate...your reference gene set can save significant plate space and may be worth the initial effort of setting it...

  39. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    Published
    Jan. 6, 2017, 3:56 p.m.
    ...exclusively activated by the small molecule clozapine N-oxide (CNO) (Krashes et al., 2011). Viruses expressing...These viruses have been distributed in over 200 packages to more than 20 countries. With this initial success...

  40. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    Published
    Sept. 22, 2015, 2:30 p.m.
    ...): 440-448. Pubmed PMID: 25849130. 2. Hata, Aaron N., et al. "Tumor cells can follow distinct evolutionary...their own work. She says, “I spend a lot of time packing, shipping, and sharing protocols one by one,” and...
Showing: 41 - 80 of 114 results