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We narrowed to 114 results for: N-pac

Showing: 41 - 60 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...
Showing: 41 - 60 of 114 results