...inducible CRISPR inhibition (CRISPRi). A) Lentiviral vector design. B) Strategy for leak-free transcriptional...highlights from the past couple months: A new lentiviral CRISPR library from Alec Kimmelman targets mouse...the new selection of ready-to-use AAV9 and AAVrg vectors. New red-shifted INTRSECT 2.0 calcium sensor available...your imaging experiments with the Cre-OFF/FLP-ON vector 137128-AAV8: Ef1a-Coff/Fon-sRGECO ...

...A newly established all-in-one vector construction system for CRISPR/Cas9-mediated multiplex genome engineering...systematically established way of making an all-in-one vector for multiplex genome engineering.” Now, they’...ease and efficiency for all-in-one CRISPR/Cas9 vectors expressing up to seven guide RNAs (gRNAs). Sakuma...Schematic overview of the all-in-one CRISPR/Cas9 vector construction system for multiplex genome engineering...expression cassettes of the gRNAs into a single vector in tandem. They went on to show that their CRISPR...CRISPR/Cas9-nuclease and CRISPR/Cas9-nickase vectors work successfully to target multiple sites in cultured...it has not yet been demonstrated, they say the vectors are also likely to work in animal embryos based...

...methylation and activate transcription. A lentiviral vector with the same modifier, Fuw-dCas9-Tet1CD, ...Core, as well as pLV-dCas9-p300-P2A-PuroR for lentiviral expression. Figure 1: dCas9-p300 adds ...Tet-dependent (TetO-dCas9-D3A) constructs. For lentiviral expression, Fuw-dCas9-Dnmt3a and Fuw-dCas9-Dnmt3a-P2A-tagBFP...hUbC-dCas9-MQ1(Q147L)-EGFP is also available for lentiviral transduction. Lysine-specific Demethylase 1 (... targeted enhancers. Although the lab used this vector in chick embryos, it also functions in mammalian...and this fusion allows for packaging into viral vectors and targeted delivery into hard-to-manipulate cell...targeting to a wider set of genomic targets. These two vectors are part of the larger pMVP/pMAGIC cloning system...

...Multiple Lentiviral Expression Systems (MuLE) Kit: This kit can be used to create lentiviral vectors expressing...into distinct expression vectors using the enzyme BbsI. These expression vectors each contain Type IIS restriction...Cas9 or dCas9-containing destination vectors. Destination vector options include humanized wt Cas9, dCas9...wt hCas9 entry vectors are supplied with the kit, you can clone your own entry vectors containing other...containing vector using Golden Gate assembly, and up to 8 gRNAs may be expressed simultaneously. Vectors for ...Cas9-based genome engineering from a single lentiviral vector.” Nucleic Acids Research 42(19) (2014): e147...assembly of multiple components into a destination vector. The first step in CRISPR/Cas9 Golden Gate multiplexing...

...Rosa26-Cas9) hematopoietic stem cells with a lentiviral vector containing a gRNA. These transduced stem cells...which utilizes custom mouse strains and lentiviral expression vectors (Table 1). The four systems Name...pXPR_071, a plasmid that leads to higher titer lentivirus preparations than the first generation CHIME ...

...categories (e.g., for a lentiviral vector to be used in mammalian cells, check Lentiviral and Mammalian Expression...the vector backbone you cloned your insert into and fill out the appropriate boxes with the vector backbone...backbone manufacturer, vector backbone size, total size of your vector + insert, any modifications you ...provide any full or partial plasmid sequences and any vector maps. The more sequence data available, the better... was a part of the insert (when cloned into the vector backbone). If your plasmid has more than one insert...you made to the backbone, and the vector type. Pro Tip! Reference the Addgene ID in the "Backbone Manufacturer...enter "Feng Zhang (Addgene plasmid # 52961)".   The vector type should describe how your plasmid is intended...

...-293T cells. If you titrate your viral preps, lentiviral titration requires an additional couple of days...require cloning specific AAV or lentiviral machinery into a baculovirus plasmid. Production of baculovirus...baculovirus construct, called a bacmid. Like AAV and lentiviral packaging plasmids, bacmids are readily available...references More resources on the Addgene blog Viral Vectors 101: Producing Your rAAV AAV Titers: Where Do They...platform to produce recombinant proteins and viral vectors. Frontiers in Bioengineering and Biotechnology,...factory to produce adeno-associated virus type 2 vectors. Human Gene Therapy, 13(16), 1935–43. https://doi.org...cells Viruses AAV and lentivirus Baculovirus; can produce AAV and lentivirus if needed # of plasmids...

...depression. The researchers found that injecting lentiviral vectors expressing BDNF in the hippocampus causes...studies using viral vectors in behavioral neuroscience. With the ability of viral vectors to fine tune gene...all of our viral vectors blog posts Find tips for getting started with viral vectors for behavioral neuroscience...is activating, inhibiting, or modulating. Viral vectors, in contrast, are considerably more versatile and...behavioral tests, but now we’ll focus on how viral vectors have changed the scope and power of three commonly... several recombinase-expressing ready-made AAV vectors which can be used in numerous floxed animal species...the availability of transgenic animals and viral vectors, scientists are combining genetic manipulations...

...display (FingRs), Xue Han’s lab has developed AAV vectors to selectively visualize excitatory and inhibitory...fluorescence from other regions of the cell. By combining vectors expressing different fluorescent proteins targeting... The Human GlycoGene CRISPR Library is a new lentiviral CRISPR library that targets human glycosyltransferase...inhibition pooled library is now available as lentiviral preps in addition to the pooled library format...library inhibits over 18,000 human genes. Find the lentiviral preps at Addgene hSyn1-SIO-stGtACR1-FusionRed...

...biosensor affinity for calcium.”  Primary Vector Type The vector type should describe how your plasmid ...indicate type in “Vector Type Other” column. For example, you may enter “donor template vector”. Cloning...: mammalian expression, bacterial expression, lentiviral, retroviral, AAV, RNAi, luciferase, cre/lox, ... (prone to recombination in bacteria) such as lentiviral, retroviral, and AAV plasmids, choose the NEB...plasmids containing the ccdB gene, such as Gateway vectors, choose the ccdB Survival strain. For plasmids ...

...the Day Lab, who developed neuron-optimized lentiviral vectors for in vitro and in vivo CRISPRa (Savell ...physiology. The Day Lab’s neuron-optimized lentiviral vectors can be found here. High-throughput screening...Neuron-optimized viral vectors Alternatively, Cas9 activators can be delivered to the CNS by lentivirus, which has.... This dual lentivirus system pairs ubiquitous expression of sgRNA(s) and mCherry (vector 1) with neuron-specific...including transgenic mice, neuron-optimized viral vectors, and high-throughput screening approaches. Here...largely precludes delivery by adeno-associated vectors (AAV). This issue was recently circumvented by ...BioRender.com. The Day Lab used their optimized vectors to highlight a key advantage of CRISPRa over traditional...

...treatment. The library consists of a pool of lentiviral vectors with roughly 73 million semi-random, 30 bp...individually label millions of mammalian cells through lentiviral infection and to monitor their abundance and ...optimize protocols for in vivo screening with lentiviral pooled shRNA libraries. Later, Bhang says, “When... Find the ClonTracer Library at Addgene Find Lentiviral Plasmids at Addgene Check out Our Cancer Resources...infection. When using the ClonTracer system, lentiviruses containing these barcodes infect the target ...

... and other viral vector tools in the years to come. We’ll make many more viral vector tools and resources... Service Page Browse Additional Viral Vectors Find Viral Vector Protocols ... and requests are still pouring in! While our lentiviral service is going strong, the AAV service has ... retrograde (the most popular form), this viral vector is used to deliver DNA encoding Cre recombinase...be sure to check back often! To find more viral vectors available through Addgene, be sure to check out...out our viral service page and browse our viral vector blog posts for many more tips and ideas on how to...thanks to Hongkui Zeng’s lab for providing this AAV vector! Additional Resources AAV Titers: Where do they...

...Addgene.org Browse Our Viral Vector Plasmids Read Our Lentiviral Vector Guide Read Our AAV Guide  ...of producing infectious virus (similar to lentiviral vectors) because they do not encode all of the necessary...on the Addgene Blog Read Other Posts on Viral Vectors Check out Our Scientist's Guide to Ebola Contribute... viral vector...

...pandemic, the Krogan lab at UCSF generated lentiviral vectors expressing SARS-CoV-2 open reading frames...which all but one are cloned into pLVX vectors (for lentiviral packaging) and contain an IRES-Puro marker...(2020) Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization...published a protocol and reagents for pseudotyping lentivirus with the SARS-CoV-2 spike protein. They deposited...

...A key feature of this system is a lentiviral gRNA expression vector (pRDA_526) that (1) enables efficient...model in vivo. In this screen, we used the pRDA_526 vector to knock out 30 genes putatively involved in CD8...robustness of the system and utility of the pRDA_526 vector. Moreover, the screen identified putative novel...

... existing transposon and vector systems and creating an all-synthetic vector that included only the elements... into a destination vector in a predefined order. Finally, the destination vectors are used with Multisite... eight entry vectors can be used for each Multisite Gateway compatible destination vector for a maximum...deposited to Addgene, including lentiviral and AAV expression vectors for certain variants. Klapoetke...step, compatible Level 0 vectors are directionally assembled into a Level 1 vector creating a single transcriptional...of interest into the pCoofy vectors, select the appropriate pair of vector and gene primers from Table...homology between the vector and gene primer for SLIC cloning. Amplify the pCoofy vector and the sequence ...

...fusion protein. Mammalian vectors One of our most popular mammalian expression vectors, c-Flag pcDNA3, comes...will need accessory packaging vectors to generate the virus. Bacterial vectors This system is great for overexpressing...cases bacterial expression vectors are inducible (e.g. pBAD LIC cloning vector (8A)) and have an epitope...folding assays. Yeast vectors There are several types of yeast expression vectors. Consider your applications...shuttle vectors. You may prefer the more traditional approach of an autonomously replicating vector that ...conduct your custom experiment. Fly vectors The vast majority of fly vectors are based on p-element, random... to reporter vectors (e.g. pBV-Luc) and gene silencing (e.g. pLKO.1 - TRC cloning vector). Check out our...

...with GeneWeld vectors GeneWeld from Jeffrey Essner’s lab uses CRISPR/Cas9 and a donor vector to create knock-ins... tools for your research! This time: GeneWeld vectors to create knock-ins CasX Drug inducible CRISPR/...cargo (ex: fluorescent protein gene) on the donor vector. Therefore, targeting requires two sgRNAS: one ...site and a universal sgRNA that targets the donor vector sequence. Homology arms that flank the donor plasmid... Read the paper in bioRxiv Find the GeneWeld vector kit CasX, a small Cas protein, is active in E....CRISPR-Cpf1(Cas12a) Double Knockout Library is a lentiviral library targeting 26 genes chosen from an analysis...

...create a final lentiviral vector containing up to 5 different elements. Lentiviral vectors constructed with...science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector backbone with ...expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT vector ensures...custom-cloned vectors in a Multisite Gateway reaction with a desired destination vector expressing a separate...combinations of genetic elements in a single lentiviral vector provides a powerful tool to modify mammalian...routes in various conditions. The plasmids and lentiviral vectors encoding these proteins can also be used ...MXS-chaining kit contains 5 empty chaining vectors plus a destination vector and a collection of building blocks...