Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 801 results for: Ran

Showing: 301 - 320 of 801 results

  1. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...protease allows a transcription factor to enter the nucleus and modulate transcription of the mCitrine ...HeLa cells were transfected with plasmids encoding FPs tagged with the plasma membrane targetin sequence...non-specific labeling. For labeling the plasma membrane, they tested different target sequences for improved...expression can vary significantly depending on transgene design and delivery. To overcome these variations...into genomic DNA with the help of the piggyBa transposase. Image from Hazelbaker et al., 2020. The lab...activating expression of TCF4 while monitoring transcript and protein levels. Plasmids for multiplexed ... new serotype PHP.V1 that exhibits efficient transduction of vesicular brain cells. Find this AAV at ...

  2. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    Published
    April 16, 2020, 1 p.m.
    ...amplification) or reverse transcriptase (RT)-RPA, respectively. RPA is coupled with T7 transcription to convert amplified... method termed DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR) (Chen et al., 2018). Both SHERLOCK...almost anywhere as lateral flow strips are easy to transport and work rapidly, providing reliable results in...    DETECTR- DNA Endonuclease Targeted CRISPR Trans Reporter Cas12a (Cpf1) is a CRISPR Cas variant that...discovered Cas12a’s ability to cleave non-specific (trans) ssDNA and harnessed this ability to create a DNA...DNase activity of Cas12a is initiated. Surrounding trans-ssDNA, including the ssDNA-FQ reporter are subsequently...Servellita V, Singh J, Miao X, Streithorst JA, Granados A, Sotomayor-Gonzalez A, Zorn K, Gopez A, Hsu ...

  3. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ...promoter of another gene, thereby driving aberrant transcription. That being said, we generally pick guides...been significant work on constructing Cas9 transcriptional activators. These constructs allow for the ...Cas9 deficient in DNA cutting activity to a transcriptional activation domain (Fig 1). Two ways to use...picking whichever guides are closest to the transcription start site (TSS). We recommend targeting the...Latency Reversal Using CRISPR/Cas9-Derived Transcriptional Activator Systems. PLoS ONE 11:e0158294 . https...Church GM (2015) Highly efficient Cas9-mediated transcriptional programming. Nat Methods 12:326–328 . https...Nishimasu H, Nureki O, Zhang F (2014) Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex...

  4. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

    Type
    Blog Post
    Published
    Nov. 27, 2019, 2:03 p.m.
    ... cell types. Specific transgene expression can be also achieved in nontransgenic animals by using cis ...delivery by increasing transgene cargo capacity, enhancing anterograde transport efficiency, and engineering...proteins that determine the AAV serotype. During transduction, the viral capsid proteins bind to the target...serotype used for a specific cell type. Find some transduction characteristics of select AAV serotypes in Table... AAV vectors are common approaches to control transgene expression by restricting expression to genetically...enhancers, multicistronic vectors, and post-translational regulatory elements. Examples of these approaches...

  5. Fluorescent Proteins 101: Fluorescent Biosensors

    Type
    Blog Post
    Published
    May 18, 2017, 2:30 p.m.
    ...enzyme or transcriptional activation, conformational changes, translation, and protein translocation. Targeting...triggers CaM/M13 interaction that increases FRET transfer between the fluorescent proteins. FRET is highly...characteristics like pH, redox state, and voltage across membranes. These sensors generally follow the same pattern...an organelle-specific targeting signal or a transmembrane domain, biosensors can be targeted to a particular...endoplasmic reticulum (ER), vacuole or cellular membranes. Examples include blue flame mitochondria-targeted...

  6. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    Published
    June 3, 2021, 1:15 p.m.
    ... the CAR facilitates signal transduction and release of cytotoxic granules from the T cell. When a CAR...., 2012, Bates and Power., 2019): rapid blood clearance, which is useful for imaging applications; better...small size (Bates and Power., 2019). Their rapid clearance from blood can be a drawback for therapeutic applications... has led to their application for basic and translational research.  Basic research applications of scFvs...single molecule tracking in living cells. The HA frankenbody is an HA tag detection probe. It contains an ...to a fluorescent protein such as GFP. The HA frankenbody works just like an antibody-based probe, but ...generate since it’s genetically encoded.   Translational applications of scFvs: Just like antibodies...

  7. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    Published
    Sept. 21, 2021, 1:15 p.m.
    ...ready-to-use viral preps   Improved cell-free transcription/translation system to examine biosynthetic pathways... Gabrielle Clouse Recently, cell-free transcription/translation systems (also referred to as TX-TL systems...modifications to edit DNA without requiring a double-stranded break. The Xiao Wang Lab has recently deposited.... These plasmids use “Cas9-mediated adenosine transient reporter for editing enrichment”, or XMAS-TREE...and enrichment use of XMAS-TREE. a) Cells are transfected with three different vectors and flow cytometry...identification of essential MYC binding sites in a wide range of human cells.  A new pooled library from Junjie...

  8. 5 Uncommon Ways to Gain Career Skills During an Uncommon Time

    Type
    Blog Post
    Published
    Oct. 20, 2020, 1:15 p.m.
    ... and not in academia is run by grant funding. Learning to write grants quickly, efficiently and effectively...apply to or offer to help with lab grant writing. You may not get the grant or your PI may edit everything...thinking about future career options. 5. Write a grant - even if you don’t need one A lot of science in...valuable skill. There are many different kinds of grants that you can apply for. If you are in a lab, your...Here are some sites that have compiled lists of grants: Science: Where to Search for Funding The Ohio...For Your Research University of Florida: Training Grants and Other Funding Opportunities L’oreal USA For...

  9. Safe Port in a Storm...How Addgene is Weathering the Pandemic

    Type
    Blog Post
    Published
    Sept. 17, 2020, 1:15 p.m.
    ...grateful to the grant officers at Fast Grants. We received a miraculous grant from Fast Grants, which was ...studying the coronavirus. Addgene’s Technology Transfer Team also moved fast to ensure that the majority...collaborations as well as provides visibility for grant-driven research being done in all labs sharing materials...materials via Addgene. Perhaps if each of many grantors supporting research could provide Addgene with... for supporting Addgene by providing a generous grant. This support will enable us to continue all operations...

  10. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    Published
    Jan. 24, 2019, 2:24 p.m.
    ...fluorescent protein (XFP) genes in a transgene. When multiple copies of the transgene cassette are introduced, stochasticity...fluorescent proteins is only possible at low copy number ranges, resulting in low fluorescent protein level. The...gene delivery is more common than conventional transgenic strategies. In such vector-based gene delivery...in the original method, we used the Tetracycline trans-activator system to boost the expression levels,...levels, and we named the system Tetbow (Tetracycline trans-activator Brainbow) (Sakaguchi et al., 2018) (Figure...expression level of XFPs, most likely by suppressing transcription. It is critical to express a minimal amount ...Bio-Protocol 8 (2018): e3046. Livet, Jean, et al. "Transgenic strategies for combinatorial expression of fluorescent...

  11. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...method is transfection of short interfering RNA (siRNA). Knockdown caused by siRNA is also transient, and ...interest. This approach often involves the transient transfection of expression plasmids in a given cell ...commercially available. Various proteomic and transcriptomic datasets can be used to select a candidate ...antibody should be able to detect even in cells transfected with low efficacy. But since there can be large...differences between endogenous expression and transfected overexpression, this approach cannot confirm ...workshops. Let’s use TIM-1 (Single pass type-1 membrane protein), which has been designated CD365 in the...the antibodies both recognized the target when transiently overexpressed in CHO cells and displayed similar...

  12. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    Published
    Sept. 6, 2017, 1:01 p.m.
    ...covalently joined, scarless DNA – perfect for transforma­­­tion or as a PCR template. Some of the applications...by using NEBuilder which requires only a single-stranded synthetic oligo to generate a single-guide RNA...RNA (sgRNA). Using only one single-stranded oligo, instead of the more typical two oligos, is more cost...annealing step usually used to create a double-stranded oligo. Generating an sgRNA construct becomes as...target region. Assembled plasmids can be directly transformed into E.coli for plating, screening and generation...complexity, is achieved with synthetic oligos with a randomized region between the regions of homology. Producing..., generating expression cassettes for yeast transformation can take 5 days. NEBuilder shortens that time...

  13. Cultivating Community Science at BosLab

    Type
    Blog Post
    Published
    Nov. 2, 2017, 1:35 p.m.
    ...2015, there were six of us. We were initially strangers but we all shared a passion for DIY Biology and...December 2015 we held our first intro to bacterial transformation course (read our blog post on it and to see...consisting of over twenty members and volunteers. Transforming citizens into scientists Anyone can become a...dedicated time at BosLab, he became our bacterial transformation guru and instructor for our intro to biohacking...biohacking courses. Our members pursue a diverse range of projects at BosLab. Some are entrepreneurs using...RNA with small molecules. Other members’ projects range from hacking probiotics to filming educational lab...BosLab. Once he optimizes plant cell culture and transgenesis he wants to share his knowledge with the community...

  14. What Good Citizenship Can Do for Reproducibility in Science

    Type
    Blog Post
    Published
    Nov. 7, 2019, 2:18 p.m.
    ...differently. Communication comes in many forms—from transparency about data to allow others to see what you actually...communication and sharing fits together to make for a transparent and collaborative research environment. Journals...Currently, most of the burden for pushing for more transparency in science communication falls on the shoulders...developed STAR Methods to improve the structure and transparency of methods sections, including clear information...researchers are not strongly rewarded for being transparent in their communication or making their data and... to a large extent, hold the keys to getting transparency and communication further up the collective ...management plans and for evidence of sharing for grant renewal—but there is clearly room to do more. If...

  15. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    Published
    Jan. 25, 2024, 2:15 p.m.
    ...Cas enzymes) cleaves both strands of its dsDNA target in cis and has no trans activity. AsCas12a, on the...cleaves ssDNA in trans. In sum, CasPEDIA documents the target substrate (DNA/RNA and strandedness) along with...Cas enzyme: Nuclease properties (CasID): Cis/trans activity, PAM requirement, gRNA, and multiplex ability...features: CasID CasPEDIA will display both the cis and trans activity of each entry. For example, SpyCas9a (one...can manually enter details to refine your search (trans activity, multiplex ability, gRNA design, etc.).....32942/X2C31F     Makarova, K. S., Wolf, Y. I., Iranzo, J., et al. (2019). Evolutionary classification...

  16. Plasmids 101: Blue-white Screening

    Type
    Blog Post
    Published
    June 4, 2015, 1:03 p.m.
    ...lactose analogue, IPTG (isopropyl β-D-1-thiogalactopyranoside). (To be completely accurate, IPTG binds...x-gal (5-bromo-4-chloro-3-indolyl-β-D-galacto-pyranoside) into galactose and an insoluble blue pigment...α-peptide) to a lacZΔM15-mutant bacterial cell in trans complements the mutation allowing for a functional...site (MCS) into the α-peptide (represented as an orange wedge in the figure on the left) and inserted it...produces pigment from x-gal (included in the transformation plate medium), turning the bacterial colony...IPTG is also included in the medium to ensure transcription of the lac operon.   Tips for blue-white screening...screening Use a good control: Transform the backbone plasmid without insert. All colonies on this plate...

  17. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    Published
    July 24, 2018, 12:51 p.m.
    ...split-vector systems’ by separating viral genes and transgene expression across several plasmids. The most commonly...separate envelope and packaging plasmids as well as transfer plasmids. This concept ensures safe handling and...Gamma-retroviral vectors have a relatively high transduction efficiency, but in contrast to lentiviral vectors... are very versatile tools as they are able to transduce dividing and non-dividing cells. More detailed...Gamma-retroviral vectors Popular gamma-Retroviral Transfer Vectors   Expressing your gene of interest pMKO...events and is widely used in the field of mouse transgenics. Experimental uses include: Cre-dependent gene... lab. Learn more about CRISPR plasmids for transcriptional inhibition. Retroviral Plasmid Kit Retroviral...

  18. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    Published
    May 22, 2018, 1:38 p.m.
    ...students first transform the S. cerevisiae met deletion strains and then isolate transformed strains on media...organism into overexpression plasmids. Students next transform deletion strains from the reference organisms ...to a whole genome duplication and extensive rearrangements in the S. cerevisiae lineage (7). In our class...class research project, students transform S. cerevisiae met deletion strains with overexpression plasmids...expression of the plasmid-encoded MET gene allows the transformed strain to grow in the absence of methionine (... to select yeast that have been successfully transformed with the plasmids, because the S. cerevisiae ...of MET/Met genes from the plasmids allows the transformed strains to grow in the absence of methionine....

  19. Streaking for Single Colonies: The Streak Plate Challenge

    Type
    Blog Post
    Published
    March 2, 2021, 2:15 p.m.
    ...and spread over the second quadrant. The process continues until all quadrants are used, hopefully revealing...students transform them into different strains of E. coli,” says Westenberg. During the transformation experiment...plate, first the bacteria are inoculated over a quadrant of an agar plate. Then, a stick is dragged through...revealing single, isolated colonies in the last quadrant after incubation. The challenge is usually successful...genetic backgrounds of the host strains affect transformation efficiency.  Aside from the Streak Plate Challenge...

  20. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    Published
    Sept. 7, 2017, 12:17 p.m.
    ...growth rate/transfection efficiency is not known, this is easily achieved by splitting transfected cells into...specific cell line, and its transfection efficiency. We often trypsinize the transfected cells on the day after...after transfection into plates containing selection agent at different densities, then dispose of the plates...under-expressing clones. If GFP-encoding cDNA is co-transfected at a favorable ratio to the gene of interest ...such as the Viaflo from Integra Biosciences) to transition from a 48-well cell plate to a 96-well assay ...expression characteristics from a population of transfected clones. The technique is applicable to a wide...
Showing: 301 - 320 of 801 results