...That’s where AAV Packaged on Request comes in.  AAV Packaged on Request With Packaged on Request, you ...AAV Packaged on Request, an expansion of our viral vectors service, has officially launched! As of today...cargo plasmids in the Addgene repository to be packaged in your choice of five different serotypes. ...following along, you may want to go to our new AAV Packaged on Request webpage to get started with your browsing...you can find your plasmids and your viral packaging service in the same place, making it easier to go from...from selection to viral vector. To start the AAV packaged on request process, simply navigate to your plasmid...plasmid of interest. If it’s eligible for packaged on request, you’ll see a banner below the ordering ...

...transfections for AAV packaging, considering using this PEI Calculator. AAV packaging typically requires ...making AAV packaging a little easier! Click here to download the PEI calculator for AAV packaging Use the...for AAV packaging Select the plasmid molar ratio you would like to use for your AAV packaging transfection...calculator will make it easier to plan your AAV packaging experiments. Let us know what you think about ...in Mammals Resources on Addgene.org Find AAV Packaging and Transfer Plasmids Here New to Making AAV? ... aav packaging...

...consider and quantify the impact of scientific publications. Ventures like Impact Story seek to find ways...holiday season want the work you are doing to have an impact, to move science forward, or perhaps even society...and has been to publish in journals with a high “impact factor,” a measure that dates back to 1975 and ... of other ways to define and measure scientific impact. Even if we stick to citation counts for a moment...October entitled “Rise of the Rest: The Growing Impact of Non-Elite Journals” found that the proportion...half.   The effect of online access on scientific impact Partly, the story has to do with online accessibility...journal publications really the best way to measure impact? As that Nature report also noted, papers detailing...

...experiment:  the packaging plasmid which contains the AAV structural and packaging genes, the adenoviral...on the packaging plasmid and take a look at each of its parts. Key elements of the AAV packaging plasmid...trans, freeing up space in the viral genome so that a gene of interest can be packaged into the virus instead...plasmid The packaging plasmid aka the RepCap plasmid encodes two genes: the replication (rep) gene and the...are required for viral genome replication and packaging: Rep78, Rep68, Rep52, and Rep40. Rep gene expression...Rep proteins from the AAV2 serotype are used to package vector genomes flanked by AAV2 ITRs into a capsid...AAV2. This production strategy is called cross-packaging.  Capsid (cap) gene The cap gene encodes three...

...tropisms.  Packaging using helper-free packaging cell lines This method utilizes a packaging cell line ... systems.  Packaging using 293T cells This method is very similar to lentiviral packaging methods. The...the viral packaging cell line. For example, Phoenix, a second generation γ-retrovirus packaging cell line...isoforms of these packaging components. Therefore, lentiviruses may not be efficiently packaged by γ-retroviral...). Cloning capacity between the LTRs is ∼8.5 kb, but inserts bigger than ∼3 kb are packaged less efficiently...has prompted the development of multiple virus packaging methods. Across these methods, the following components...) or MSCV (Murine Stem Cell Virus) sequences. Packaging genes (viral Gag-Pol): Gag is a structural precursor...

...ground-breaking technology that’s already had a huge impact on research and will be affecting our everyday ...with engineered variants: eSpCas9, SpCas9-HF1, & HypaCas9. The off-target problem It is well known that... dynamic structure leads to the development of HypaCas9 Researchers in the Doudna Lab at Berkeley recently...recently added their own hyper accurate Cas9 variant (HypaCas9) to the pool of enhanced CRISPR tools. In their... Lab, they used targeted mutagenesis to create HypaCas9 (SpCas9 N692A/M694A/Q695A/H698A). This new variant...when compared to eSpCas9(1.1) and SpCas9-HF1. The HypaCas9 plasmids are available from Addgene here! Future...enhanced specificity, eSpCas9, SpCas9-HF1, and HypaCas9 should enable researchers to make precise edits...

...vector, but its packaging capacity is limited to ~4.7kb, or roughly half the packaging limits of lentiviral... up packaging an oversized genome of ~9 kb into AAV. They found that in defiance of the packaging limit...2016) Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids. Human ...System Expands Adeno-associated Viral Vector Packaging Capacity in a Transgene-independent Manner. Molecular...determined that even when larger vector genomes are packaged, the physical size of the genomes is still ~4.7kb...oversized AAV genomes are fragmented or truncated when packaged and then following transduction, the partial genomes.... Each piece of the gene is then independently packaged into an AAV. When a cell is co-infected with both...

...known PID formats are resolvable compact identifiers, which are Compact Uniform Resource Identifiers (CURIEs...the specific plasmids used. RRIDs and resolvable compact identifiers now exist for all Addgene plasmids!... Addgene now provides an RRID and a resolvable compact identifier for every plasmid. In case these PID...which has the Addgene ID #87155. The resolvable compact identifier format for lenti-Cas9-VQR is the URL...to include the option of citing with resolvable compact identifier and RRID formats. Let’s go back to our...the entire line. Cite plasmid using a resolvable compact identifier and RRID “lenti-Cas9-VQR-Blast was a...Addgene_87155)” Cite plasmid using a resolvable compact identifier “lenti-Cas9-VQR-Blast was a gift from...

...actions     Use active listening skills such as the PAC Method. P = demonstrate patience, pause; A = ask ...

...distribution and relevance for the efficiency of BAC/PAC recombineering techniques. Semprini S, Troup TJ, ...

...inhibits chain elongation. 50-500 ug/mL Puromycin pac HeLa, 293HEK, Jurkat T cells Inhibits protein synthesis...

...over to our CRISPR Plasmids and Resources page.  PAC-MAN (prophylactic antiviral CRISPR in human cells...

...effects. The library was cloned into the Liu lab’s own pAc-sgRNA-Cas9 insect expression backbone, which expresses...be found here). To use the library, the authors packaged it into lentivirus and used the resulting lentiviral...large size of dCas9-VPR limits one’s ability to package it into lentivirus and later create stable cell...cells. The piggybac system has a much larger cargo capacity than lentivirus and is capable of efficiently ...

...1,000 active makerspaces around the world. Hackerspaces.org maintains a list of active spaces near you. ...@derekja  Resources Hackerspace Website https://wiki.hackerspaces.org/Hackerspaces OpenWorm Project Website...manifested itself in the creation of Hackerspaces and Makerspaces back in 2007, which function as centres...with the OpenWorm Project One such space is the Victoria Makerspace in Canada, which I belong to. The ...  Funding Makerspaces One of the main challenges when starting a new project at a makerspace is finding... can find a makerspace somewhere near you. The place to start looking for your local space is the Hackerspace...competition. In 2010, he founded makerspace.ca, Victoria’s first hackerspace which now offers members and ...

... take a look at the RSpace ELN.  The connected approach in action: how the RSpace ELN supports reproducibility...repositories. RSpace has been integrated with two of the five: Figshare and Dataverse, as well as the DSpace repository... reproduce an experiment documented in RSpace using the RSpace datasets deposited in Figshare or Dataverse...Rory is founder and CEO of Research Space, which provides the RSpace electronic lab notebook. Rory is based...contributed by Rory Macneil, founder of Research Space. There are many types of electronic lab notebooks...reproducibility RSpace was designed in response to requirements that emerged from intensive engagement with...byproduct of connectivity. Between 2012 and 2017 RSpace was developed with these principles in mind, and...

...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...reduce the risk of such events, newer lentiviral packaging systems divide the viral elements among multiple...

...lentiviruses the same thing? Can I package lentivirus with retroviral packaging plasmids (and vice versa)? A1...plasmids can be packaged with either system. Please note that the generation used to package the virus does...co-transfecting three essential ingredients: the lentiviral packaging vector, a transfer vector including the gene of... an envelope. Three generations of lentiviral packaging systems have been developed over the years based...most experiments; however, the 3rd-generation packaging systems offer the maximal biosafety available ...different plasmids into the producer cells (two packaging plasmids, an envelope plasmid, and the lentiviral...and gamma-retroviruses use the same genes for packaging (that's gag, pol, and env), the isoforms of these...

...receive your Packaged on Request prep 6–9 weeks after placing your order.  Can I order Packaged on Request...expanding our viral vector service to include a Packaged on Request option. Through this expansion, scientists...Figure 1: The process of requesting and ordering a Packaged on Request viral vector prep.    Excited? ...in the Addgene repository will be available for Packaged on Request service. A small subset of AAV plasmids...launch, we will offer five different serotypes for packaging: AAV1, AAV2, AAV5, AAV8, or AAV Retrograde (AAVrg...expansion happen?  You will be able to request Packaged on Request service starting in January 2025.  ...requestors.  Requesting Plasmids available for Packaged on Request consideration will have a banner on...

...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...virus via recombination between the transfer and packaging plasmids does exist. To further reduce this possibility...