...packaging plasmids. The cells need three different plasmids to produce AAV: 1) the RepCap plasmid which provides...AAV plasmid standard and AAV transfer plasmids. It’s easy to contaminate a workspace with plasmid, not... viral replication; and 3) the transfer plasmid which encodes a transgene of interest that’s packaged ...going to lie, AAV transfer plasmids are kind of a pain to work with. These plasmids contain ITRs which are...cultures of your AAV transfer plasmid at 30 °C instead of 37 °C and then screen for ITR recombination with...to miniprep 5 mL of my AAV transfer plasmid cultures and then screen with SmaI digests. I freeze the bacterial...AAV’s protein shell, or capsid; 2) the pHelper plasmid which expresses adenovirus genes which help mediate...

...μg pLKO.1 shRNA plasmid 750 ng psPAX2 packaging plasmid 250 ng pMD2.G envelope plasmid to 20 μl serum-...to vary the ratio of shRNA plasmid, packaging plasmid, and envelope plasmid to obtain the ratio that gives...shRNA. psPAX2 Packaging plasmid for producing viral particles. pMD2.G Envelope plasmid for producing viral... also be used with packaging plasmid pCMV-dR8.2 dvpr and envelope plasmid pCMV-VSVG from Robert Weinberg...yield of DNA. Back to Top D. Screening for Inserts You may screen for plasmids that were successfully ligated....1 Protocol pLKO.1 - TRC Cloning Vector Addgene Plasmid 10878. Protocol Version 1.0. December 2006. Copyright... RNAi Consortium A.2 Map of pLKO.1 A.3 Related plasmids B. Designing shRNA Oligos for pLKO.1 B.1 Determine...

...received over 13,000 plasmids requests for COVID-19 related plasmids. These plasmids have reached labs in...small collection of plasmids for coronavirus research, including a handful of plasmids that have been in...some highlights of the plasmids shared over the last year and how these plasmids have been used to accelerate...circulating and plasmids containing these mutations have been deposited at Addgene. Plasmids deposited from... labs also deposited plasmids for producing COVID-19 diagnostic enzymes. Plasmids in this toolkit include...These plasmids contain both untagged and tagged versions of SARS-CoV-2 ORFs and include plasmids that ...scientists through plasmid sharing and more. As you’ve read above, some of these plasmids have been in the...

...Allen Institute for Cell Science Plasmid Collection Check Out Plasmids for FRET Check Out the Fluorescent...cloning your protein of interest into a vector encoding a fluorescent tag. Once expressed as a fluorescent... on the gene names in the figure below to find plasmids containing that particular gene. ... For subcellular markers in yeast see our plasmid collection from Sue Jaspersen (Stowers Institute...Prescott (Monash University). You can find more plasmids for labeling your subcellular structure of interest...interest in the Allen Institute for Cell Science Plasmid Collection. You can also use the library of full-length...Localization Poster here! References 1. Allen Institute Plasmid Pages 2. Cooper GM. 2000. The Cell: A Molecular...

... Plasmid Collections EXPRESs Plasmids EXtracellular Plasmid RESource (EXPRESs) Consortium... Wright Lab plasmids EXPRESs is a collection of mammalian expression plasmids that encode the extracellular...commonly-used bait and prey plasmids can be found here: Plasmid Type Plasmid Name Bait Cd4d3+4-bio Prey...antibody plasmids P. falciparum merozoite surface protein plasmids P. vivax merozoite protein plasmids Human...Consortium plasmids that encode the extracellular regions of cell surface receptor and secreted proteins...approaches have created this resource to facilitate open and free exchange of these expression plasmids. It is ...to join our community and contribute their own plasmids in the future. AVEXIS: One popular method to detect...

...Collections INTRSECT Plasmid Collection INTRSECT Plasmid Collection INTRSECT Implementation Plasmids References ...of populations expressing only Cre AND Flp, Cre NOT Flp, or Flp NOT Cre. B) This strategy has been expanded...AND NOT Cre Yes 137129 pAAV-Ef1a-Con/Fon-BFP Cre AND Flp Yes 137130 pAAV-Ef1a-Con/Foff 2.0-BFP Cre AND NOT...intersectional expression plasmids that combine multiplexed recombinases, including Cre, Flp, and VCre to enable viral...expressing Cre AND Flp, here expressed through a double transgenic animal strategy (PV-2a-Cre; SOM-IRES-Flp...novel INTSRECT plasmids for groups interested in producing their own INTRSECT plasmids. Online spreadsheet...genetically-encoded calcium indicators, and rabies targeting genes. Recombinases Addgene ID Plasmid Logic Sites...

...this evolving field! Find prime editing plasmids! Find plasmids from David Liu's lab! This post was originally...the CRISPR guide Find CRISPR plasmids by function Browse our entire plasmid collection ...target site by the pegRNA — a guide RNA that also encodes the desired edit and homology to the genomic DNA...require multiple steps and enzymes. Prime editing can encode any of the 12 possible base-to-base changes in ...for treating genetic diseases. In our summer Hot Plasmids post, we covered the Liu lab’s first demonstration..., the Liu lab created and evaluated 19 PE1 variants with RT mutations known to increase activity, enhance... initial nick site (to avoid creating a double strand break), increasing editing efficiencies 2-3-fold...

...their MECP2-related plasmid tools with Addgene and share additional resources. Plasmid tools to facilitate...to partner with them to launch a Rett Syndrome Plasmid Collection and Resource Center for Rett Syndrome...(Guy et al., 2007). They accomplished this using Cre-Lox mouse models in which the endogenous MECP2 gene...excision of the Stop cassette using tamoxifen-inducible Cre not only halted the neurological damage but completely... Rett Syndrome in novel ways.  Addgene’s MECP2 Plasmid Collection and Resource Center will continue to...syndrome is caused by mutations in X-linked MECP2, encoding methyl-CpG-binding protein 2. Nat Genet 23:185...two MECP2 labs (Gail Mandel and Stuart Cobb) to create the Gene Therapy Consortium. Leveraging their combined...

... Check out All the Plasmids from the Collin's Lab Find Synthetic Biology Plasmids for Your Next Experiment...deposited the best performing sensors and constructs encoding corresponding portions of the Zika virus genome...Components." Cell (2016). PubMed PMID: 27160350. Find Plasmids from the publication at Addgene 2. Green, Alexander...Experiment Deposit Your Plasmids with Addgene ...parallel. More broadly, we drew on the combined creativity and expertise of the group and yet never hesitated...

...transcription from the accessory plasmid (AP) encoding phage gene III. The accessory plasmid library contained all...mutagenesis plasmid (MP6) permits quick, large-scale mutation of the gene of interest. They created a selection...doi: 10.1038/nature26155 PubMed PMID: 29512652 Plasmids from this paper are available at Addgene.  Additional...Addgene.org Check out our CRISPR Guide Find CRISPR Plasmids for Mammalian Systems Learn about CRISPR History...genome-wide off-targets, since increasing PAM flexibility might also increase off-target activity. Surprisingly...gone one step further, using directed evolution to create xCas9, an enzyme recognizing a broad range of PAMs...PAMs like NG, GAA, and GAT, but also displaying increased editing specificity. We’re excited to learn more...

...Gonçalves and Ventura labs’ plasmids, including protocols, check out the plasmids at Addgene: pAdSh.PGK.Cas9...looking for a broader range of CRISPRs plasmid tools, find more plasmids, CRISPR technology guides, FAQs, and...mutagenesis similar to those achieved by isogenic AdVs encoding TALENs targeting the same chromosomal region. ...description of their experimental protocol. The three plasmids which have been deposited to Addgene are: pAdSh.PGK.Cas9...templates in somatic cells of adult animals in order to create animal models for particular cancer. That’s exactly...the Memorial Sloan Kettering Cancer Center did to create a model of Eml4–Alk-driven lung cancer (Maddalo... Addgene and could be useful to labs wishing to create this kind of animal models. Start using adenoviral...

...dTomato Activity Cre-dependent Flp-dependent Cre and Flp-dependent Constitutive (non-cre dependent) Serotype...Ready-to-use AAV available from Addgene's viral service encoding chemogenetics tools. Viral... in specific cell types. See our Chemogenetics Plasmids and Chemogenetics Guide pages for more information...Filters ID Name Receptor and Activity Reporter/Tag Cre requirement Serotype PI 44361 pAAV-hSyn-DIO-hM3D(...Gq)-mCherry hM3D(Gq) - Activation mCherry fusion Cre-dependent 1, 2, 5, 8, 9, rg*, PHPeB Roth 44362 pAAV-hSyn-DIO-hM4D...Gi)-mCherry hM4D(Gi) - Inhibition mCherry fusion Cre-dependent 1, 2, 5, 8, 9, rg*, PHPeB Roth 65417 pAAV-hSyn-dF-HA-KORD-IRES-mCitrine...kappa-opioid receptor (KOR) - Inhibition IRES mCitrine Cre-dependent 8 Roth 50454 pAAV-hSyn-DIO-HA-hM3D(Gq)-...

...available Addgene plasmids he or she can use to study it (as seen in the screenshot below). Currently...refers to 3 proteins (H-Ras, K-Ras, and N-Ras) encoded by a family of proto-oncogenes. Ras proteins are...generation of mutant cell lines. Further, for each plasmid in the kit, the transcript most commonly found ...initiative to curate our collection of cancer-related plasmids such that each is linked to a known major pathway...pathway. Now, using our new Cancer Research Plasmids and Resources page, investigators may select a pathway...Visit our Cancer Collection Pages Deposit Your Plasmids for Studying Cancer Browse Our Other Collections...as 30% of all cancers, especially those of the pancreas and colon. Despite the prevalence of Ras mutations...

... target plasmid and pX330-sgRNA plasmid (Mashiko et al. 2013). The pCAG-EGxxFP target plasmid contains...pCAG-EGXXFP plasmid along with the individual gRNAs in their PX330 plasmids (or similar plasmids for gRNA... (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences ...effective ones to increase your chances of gene modification in vivo. Find Plasmids for Testing Your gRNAs... system floating around, and many pre-designed plasmids containing these variants ready for purchase. ...multi-cloning site (MCS) between EGFP fragments. The pX330 plasmid contains humanized Cas9 expression cassette and... you can insert this region into a pCAG-EGXXFP plasmid using standard cloning techniques (The XX indicates...

... and Packaging Plasmids Common γ-retroviral envelope and packaging plasmids. ID Plasmid Type Description...envelope plasmid Kasparov Popular γ-Retroviral Transfer Plasmids Common γ-retroviral transfer plasmids. Want...retroviral plasmids at Addgene, including popular packaging, envelope, and transfer plasmids. Viral...Plasmids γ-Retroviral Plasmids γ-Retroviral Plasmids Additional Resources: γ-Retrovirus Guide Biosafety...Biosafety Guide Lentiviral Plasmids AAV Plasmids Viral Vectors 101 eBook γ-Retrovirus (gamma-retrovirus) is an...with packaging plasmid Weinberg 12259 pMD2.G Envelope VSV-G-expressing envelope plasmid Trono 1733 pCI-VSVG...envelope plasmid Nienhuis and Salmon 35616 pCAG-VSVG Envelope VSV-G-expressing envelope plasmid Nienhuis...

...beneficial for plasmid clearance. It is more convenient to update the spacer(20nt) on pEcgRNA plasmid as only.... pCutamp cures the plasmids by targeting the ampR promoter in all three plasmids. pCutamp is then removed... 13 plasmids with 14 rounds of transformation in 30 days. Figure 2: (A) MUCICAT plasmids for ... original pCas/pTargetF system consists of one plasmid that expresses the S. pyogenes Cas9, a temperature-sensitive...the pTargetF construct and then introduce both plasmids into the cell. The edit in the chromosome can ...the pMB1 replicon, resulting in cutting of the plasmid. At this point, subsequent introductions of pTargetF...addition of the sgRNA sequence in pEcgRNA. Next both plasmids are transformed in E. coli with the donor DNA ...

...s lab at UC Davis is to generate a genetically encoded fluorescent sensor for serotonin. According to ...Addgene.org Find fluorescent protein biosensors Browse Plasmids from the Tsien lab Browse the Michael Davidson...site-saturation mutagenesis, and medium-throughput screening, she increased the affinity of the sensor for serotonin...overwhelming response and were excited to see the many creative uses of fluorescent proteins. We’re happy to announce...thousands of protein variants simultaneously  With increasing sequencing power, scientists are able to quickly... protein-coding pathogenic human variants are decreased in protein abundance, possibly due to misfolding...neuromodulator, and evidence suggests that it is incredibly important for nearly every aspect of life.” Unger...

...Antibodies Resources on Addgene.org Find plasmids encoding recombinant antibodies at Addgene ...could mean needing to increase crosslinking to counteract the effects of increased sample agitation; a ...such as qPCR or next-gen sequencing. ChIP is an incredibly useful application for exploring these interactions...crosslinking, which uses formaldehyde or UV light to create strong but reversible bonds. This will keep the... necessary for most ChIP experiences, but the increased binding can mask the epitopes antibodies recognize...shorter sonication time could be an opportunity to decrease crosslinking. (Homogenization for a tissue sample...

...pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR This plasmid contains two expression cassettes, Renilla luciferase-2A-Cre recombinase...sgRNA(LacZ)-pCBh-Cre-WPRE-hGHpA-ITR This plasmid contains two expression cassettes, Cre recombinase and...(backbone)-pCBh-Cre-WPRE-hGHpA-ITR This plasmid contains two expression cassettes, Cre recombinase and...ITR-U6-sgRNA(NeuN)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR This plasmid enables Cre/loxP recombination and...sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR This plasmid facilitates Cre/loxP recombination ...tracrRNA: This plasmid contains three expression cassettes. To target a given site, the plasmid can be digested... vector encoding dCAS9-VP64 with 2A Blast resistance marker 61426 : lentiviral vector encoding the MS2...

...al., 2007). Brainbow uses a Cre/lox system to label neurons. Genes encoding the fluorescent proteins are...fluorescent protein genes, Cre recombination is required before fluorescence can occur. Cre recombinase can be...developing fish via Cre protein microinjection or crossing of Zebrabow animals to an inducible Cre line. The Schier...gene directly following the promoter is expressed. Cre-mediated deletion of one or several of these genes...system. One such system is Brainbow 2.0, which uses Cre to trigger the inversion of two genes aligned head-to-head...head-to-head. Another variation, Brainbow 2.1, uses Cre to trigger the inversion of two tandem sets of genes...a specific tissue. XFPs are introduced using a plasmid construct called Zebrabow-GateDest containing an...