Skip to main content
Addgene

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

In Category

We narrowed to 1,078 results for: CAN

Showing: 401 - 420 of 1078 results

  1. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    ...undergo nick repair. This method can work well, but poor template digestion can result in a high number of ...Cloning, and SLIC can all be used to assemble multiple fragments, but these methods can be expensive because... causation in molecular biology and genetics. It can be used to characterize the function of certain regions...biomedical research, modeling patient mutations using SDM can help determine if a variant is causal for a given...strategies, only 3-4 phage proteins are needed and these can be provided via a separate plasmid or be inserted...machinery is induced using L-arabinose. These cells can be made and frozen in large batches suitable for ...

  2. How to Lead a Great Meeting

    Type
    Blog Post
    ... to our “real” work. However, a well-run meeting can have quite the opposite effect. A great meeting should... meeting. Follow the tips below to learn how you can run a top-notch meeting. Define the meeting Define...some other commitment. An announcement of purpose can help everyone to ‘land’ in the meeting and get focused... direction you’d like to pursue? Undefined scope can lead to long meetings spent on details that do not... sure to keep them manageable. At Addgene, teams can add as many points as they like to the written agenda...we like to send around a Google doc where people can make comments. At meetings, we just focus on the ...meeting (see “defining the scope” above) or if you can cut it down to thirty minutes. Keep it moving: ...

  3. Measuring Kinase Activity at the Single-Cell Level with Kinase Translocation Reporters (KTRs)

    Type
    Blog Post
    ..., which can be difficult to predict. Slower Dephosphorylation Rates: FRET-based reporters can have slower...measuring kinase activity at the single-cell level can aid in drawing connections between signaling activity...spectral properties that limit which fluorophores can be multiplexed together. Challenging  to Design: ...cytoplasm and/or nucleus, while FRET-based reporters can be used to measure kinase activity in specific subcellular...and the cytoplasm, nuclear export and import rates can influence KTR translocation. Work from the Covert...

  4. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ...your lentiviral vector will infect, pseudotyping can help. In pseudotyping, you produce viral vectors ...cytotoxicity and the mechanisms behind the cytotoxicity can vary. For example, the popular VSV-G envelope protein...syncytias that cause the cells to die. Cytotoxicity can also be cell type dependent: if a cell has few receptors...lentivirus (Crawford et al., 2020). That way, they can study the spike protein’s role in infecting cell ...methods.  In terms of preparation, VSV-G is stable and can withstand ultracentrifugation which helps scientists...cells and neurons. However, VSV-G’s broad tropism can be undesired for in vivo studies since there’s the...the potential for off-target infectivity as it can bind and transduce many cell types before reaching ...

  5. Transferable Skills Guide: External Collaborations

    Type
    Blog Post
    ...methods and machines one team can learn so linking together expert teams can push a scientific project to...too), we will often look for partnerships where we can leverage another organization’s expertise to our ...meeting people with different mindsets and approaches can truly broaden your mind. When I first began collaborating...Getting companies to come to these types of events can be very tough. First I needed to find the right contacts...guidance. The relationships you establish in this way can, of course, lead to jobs and even favors, but you...don’t forget about collaborations and projects you can do with others after work. Get sponsors for a training...accomplish with people at your organization. You can meet collaborators  for lunch, make a point of talking...

  6. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...control of DNA recombination and gene expression can be advanced by the coordinated use of fellow SSR ...asymmetric core. Recombination between these sites can lead to excision, inversion, and translocation of...Continuous inversion of the DNA sequence of interest can lead to poor expression of genes within that sequence... for Cre and FLP function. Because recombination can only occur between target sites of the same sequence...recombinase - keeping Cre out until needed But what can be done when the recombination should only occur ... cortical neurons and their synapses.   How can you label a single neuron and its synapses in the...the living brain? Cells of a specific brain area can be transfected by in utero electroporation. To achieve...

  7. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ... DNA strands - a double-strand break (DSB). DSBs can be caused by intracellular factors such as nucleases...identical to the original DNA sequence at the DSB, or it can introduce specific mutations into the damaged DNA...displaces one strand of the homologous DNA donor and can template repair off the other strand; this results... loop (D loop). The recombination intermediates can then resolve to complete the DNA repair process ...Homology directed repair pathways HDR mechanisms can be divided into two main camps: non-conservative ...presence of only one invading strand at a DSB that can be used for repair (a one-ended DSB). This single...HDR is a powerful site-specific knock-in tool that can be harnessed by researchers for many applications...

  8. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...research as well. For example, CRISPRa can be used to find potential cancer drug targets by finding genes that...these can be difficult to construct and deliver when compared to gRNAs. Additionaly, cDNAs cannot be used... the potential to be used in therapeutics, as it can increase gene expression at certain loci to create...al., 2016). Additionally, CRISPR-based activation can be made inducible upon environmental triggers. This...This is done by creating a split dCas9 protein that can reassemble when exposed to specific UV wavelengths...under native conditions, the less activation you can achieve using a CRISPRa. Your gene of interest might... four guides directed towards that gene as there can be a large difference in guide potency. Thus, in ...

  9. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    ...2002) and many downstream complexes involved, it can be difficult for scientists to understand how GPCR...transducers (collectively known as the transducerome) that can be activated downstream of GPCRs that are not involved...by a GPCR on the cell’s surface into action that can result in the initiation of a vast number of signaling...downstream signaling pathways.  This dissociation can be measured using a technique known as Bioluminescence... a decreased GFP2 emission. The activation state can be measured by a BRET2 ratio, which is the ratio ...GFP2 emission to RLuc8 emission (see figure). What can TRUPATH be used for? The Roth lab used the TRUPATH...transducerome from different GPCRs. This collection can be used to study endogenous ligands, the pathways...

  10. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    ...plasmids can be used to do include: Produce large amounts of a protein so that scientists can purify and...one-stop reference guide for plasmids, so that you can spend less time researching the basics and spend ...point for PCR amplification or sequencing. Primers can be exploited for sequence verification of plasmids...make more of the plasmids as they grow, scientists can easily make large amounts of plasmid to manipulate... Produce proteins that glow so that scientists can track their location or quantity inside a cell ...genome engineering) Produce synthetic viruses that can be used in research or for therapeutics Addgene ...

  11. RUBY-Red Siliques

    Type
    Blog Post
    ...against CRISPR positive plants, but screening these can be challenging and requires specialized equipment...mutant plants, but the seeds needed for experiments can only be collected from heterozygous individuals. Only..., T., Sun, H. et al. Hortic Res. (2020).   I can do this by expressing a gene that rescues the mutations... (Top right, second panel) Transformed seedlings can be selected without antibiotics. (Top left) Mature...homozygous RUBY plant. (Bottom panels) RUBY expression can vary from different transformation events likely ...microscope. First, red CRISPR-positive transformants can be produced. In the following generation when CRISPR...to prevent further editing, then green offspring can be selected instead of red ones. Other innovative...

  12. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

    Type
    Blog Post
    ...and can’t be contained to a specific area. Blue light, however, is spatially confined and you can see ...in the shakerTurn the LEDs onAnd when you wantYou can just turn them offNo need for any washNo need for...cloned gene of interest, under the pBAD promoter, can be controlled with light without the need of any ...have less heterogeneity in the population and you can do all of these more spatially confined experiments...are exposed will fluoresce. With these images, you can see just how spatially precise the activation is....'s lab have created and shared with Addgene. You can find all the plasmids deposited from Di Ventura's...here. Optogenetic plasmids from the Khammash lab can also be found here.   References and Resources References...

  13. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    ...that now can be deposited at Addgene. What makes these fluorescent proteins unique and what can they be...mWasabi can be used with Sapphire (excitation maximum 399 nm) because it does not significantly excite ...green and yellow fluorescent protein wavelengths, it can be imaged either with standard green fluorescent ...reduction in emission readout, respectively. mNeonGreen can be used as a fusion tag for a wide range of applications... other common fluorescent proteins and therefore can be used in immunoprecipitation experiments with other... mCerulean imaged with a standard CFP set. mTFP1 can be used as a FRET donor to mCitrine and mOrange or...

  14. Oh, The Places You Can Go: Careers in Science Communication - Curriculum Development

    Type
    Blog Post
    ...background. [We ask them,] 'Can you read this lab. Can you test this experiment? Can you follow and understand...subject matter expert in the educational sphere, it can range from researching an area, to writing an introduction...and operations departments at eScience Labs. This can involve traveling with the sales team to meet faculty... since the eScience warehouse is onsite, Caitlin can visit the operations team to make sure “that everything...and look at new technologies in education and ask ‘Can we implement that?’ [And think about] where we [eScience... staff writer at the non-profit Fred Hutchinson Cancer Research Center. And make sure to subscribe to ...

  15. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    ...constitutive high level expression? Finally, you can add bells and whistles to your construct. If you ...specific antibody against the protein available – you can always add an epitope tag or a fusion protein. Mammalian...visualize YGOI in vivo (try pcDNA3-EGFP). If you can get away with generating transiently transfected ...selection cassette (e.g. 1435 pSG5L Flag HA). You can also upgrade your basic mammalian expression system...containing pDEST-HisMBP). After purification the protein can be released from the resin and used for various binding...Consider your applications and experimental design. You can create a new strain with a disrupted gene by transforming...under the Gal4 inducible UAS promoter. Other vectors can be expressed in fly culture cells like S2 and carry...

  16. DNA Purification Without a Kit

    Type
    Blog Post
    ...plasmid DNA sample can be treated with RNase to remove RNA contamination and you can use a phenol chloroform... filter. Each glass filter can capture up to 150 ug of DNA, and filters can be stacked together in series... equipment. Plus they create plastic waste. Kits can also have the annoying tendency to runout right when...hours to complete, while this syringe-based method can be completed in 20 to 30 minutes. Additionally, while... purposes, with the Kim and Morrison method, you can just adjust the number of glass filters you use depending...cellulose directly into a PCR buffer, where DNA or RNA can be amplified. Compared to a commercial magnetic bead-based...yields as fresh columns, suggesting that columns can be reused at least five times. DNA purification kits...

  17. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...which can “drag and drop” kilobases’ worth of DNA with CRISPR-driven precision into a genome, can be considered...collateral" activity of the enzyme can lead to off targets and result in significant, negative effects in RNA targeting...forward for what the field can do.    References and Resources References  Özcan A, Krajeski R, Ioannidi...tests samples of new bacteria from anywhere they can find, whether the samples are from a rain gutter ... instance, a disease like cystic fibrosis, which can be caused by thousands of different mutations, all...lab switched up the serine integrase for one which can enzymatically take a genetic sequence and integrate...cargo via Cas9-directed integrases.   Use PASTE can be delivered via a single dose of plasmids, works...

  18. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...issues as a region of interest can be targeted using a gRNA, which can easily be cloned into a gRNA expression... as dCas9 (Jinek et al., 2012). Like Cas9, dCas9 can bind to a specific DNA sequence via a targeting gRNA...DNA-binding molecule, such as Zinc-finger or TAL proteins, can be used. The Fujii lab demonstrated this in mammalian...proteins targeting a specific genomic locus, which can require multiple cloning steps (Fujita et al., 2013...identification of DNA binding proteins, this method can characterize nucleic acid-nucleic acid interactions...successfully used in this "in vitro" form. Any tag system can be used to isolate genomic regions of interest with...biotin ligase. Biotinylated dCas9, with bound DNA, can then be purified by streptavidin high-affinity purification...

  19. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ... language is the basis of shuttle vectors, which can function in unrelated host species, even from completely... Figure 1: Shuttle vectors are vectors that can function in more than one host species. Created with...plasmids? If we all use the same molecular code, can I transfect any old bacterial plasmid into HeLa cells...Agrobacterium tumefaciens, a plant-infectious bacteria that can incorporate foreign DNA into plant genomes. Eukaryotes... wall synthesis. Eukaryotes, with no cell walls, can grow alongside these antibiotics just fine. This ...function. A puromycin resistance gene on your vector can protect both plasmid-carrying bacteria and eukaryotes...auxotrophic cells die. Some auxotrophic selection markers can be used across species, for example, in both S. cerevisiae...

  20. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    ...spotlight — Cas3. Cas3 can induce genomic alterations on a scale that other Cas effectors can’t, while maintaining... brought to the target location using a gRNA and can then recruit Cas3, which carries out the brunt of...abundant type in the Class 1 category. These type Is can then be further broken into subtypes of A–G, which...appears to be conserved. This DNA degradation portion can be bypassed by leaving out Cas3 — a tactic some CRISPR..., developing Cas3 tools has been challenging. It can be difficult to ensure comparable expression of all...efficiency of these systems, though consistent expression can still be a challenge. A multitasking Cas Most Cas...pulled through Cas3. After these initial steps, Cas3 can perform its helicase and nuclease functions simultaneously...
Showing: 401 - 420 of 1078 results