...plant expression vectors containing several cassettes and generate multi-construct transgenic plants -...addition to individual plasmids, Addgene carries several useful toolkits and plasmid collections for use...includes plasmids containing promoters that drive general and species-specific gene expression (see the column...CRISPR Please refer to Addgene's CRISPR Guide for a general introduction to CRISPR technology or the Plant ...selection and multiplex CRISPR/Cas9 assembly for fast generation of multiple Arabidopsis mutants - Niko Geldner...MoClo Plant Parts II and Infrastructure Kit for hierarchical assembly of plant multigene constructs - Johannes...publications are created. Papers from the plant biology literature are discussed with one of the authors. The podcast...

...Expression mEmerald 487 509 39 6 Monomer (A206K) mEmerald-N1 - Mammalian Expression mEmerald-C1 - Mammalian...Expression mPA-Emerald 487 509 - Monomer (A206K) mPA-Emerald-N1 - Mammalian Expression mPA-Emerald-C1 - Mammalian...HisD-rsTagRFP - Bacterial Expression rsEmerald 491 508 rsEmerald-pBAD - Bacterial Expression rsGFP1 493...Expression Species . The content for this page was generated with the help of Erik Snapp . Empty Backbones ...Mammalian Expression mEmerald-pBAD - Bacterial Expression EGFP 488 507 34 6 Prone to dimerization pcDNA3-...

...full-text of the publication here . Article Citation Generation of Genomic Deletions in Mammalian Cell Lines ... incubation time allows for sufficient cell proliferation for screening genomic DNA (gDNA) for the intended... and 7.1 ). The bulk cells have sufficiently proliferated when the concentration exceeds ~100,000 cells...80% confluence. The clones have sufficiently proliferated once macroscopically visible with ~2 mm diameter...pellets in 50 μl of DNA extraction solution. NOTE: Generally ~100,000 cells are used for DNA extraction, although... band” in separate reactions. NOTE: Numerous polymerases may be used for step 6.3 . Run samples in a thermocycler...biallelic deletion clones with a proofreading polymerase and clone the amplicons ( e.g., with a PCR cloning...

...accessory plasmids available from Addgene for generating your own specific TALE plasmids.... TALENs assembled by Golden Gate. Validated in several species. Takashi Yamamoto Additional Plasmids for... Gate TALEN kit (Bogdanove/Voytas) in order to generate TALORs (TAL Orthongal Repressors) that can be ...Grunwald pCS2TAL3-DD and pCS2TALE3-RR are next generation TALEN backbone vectors designed for use with ...segments (136AA and 63AA, respectfully). This next generation architecture has been shown to increase mutation... C63AA) and showed robust cleavage activity in several later studies. pCAG-T7-TALEN(Sangamo)-Destination.... In addition, a T7 promoter was included for generating TALEN mRNAs through in vitro transcription as...

... regulator controlling cell growth and proliferation. mTOR, a serine/threonine kinase, is found ...mutations in negative regulators of mTORC1. The hyperactivation of mTORC1 in cancer promotes the conditions...conditions needed for uncontrolled cell growth and proliferation, including an abundant supply of macromolecules...energy, as well as the presence of both pro-proliferative and anti-apoptotic signals. mTORC2, which appears...upstream of Akt, also promotes cell survival and proliferation. mTOR Pathway Plasmids Click on a name to find...specific relationship. The content for this page was generated with the help of David Sabatini . mTOR Pathway...specific relationship. The content for this page was generated with the help of David Sabatini . Return to top...

...Institute for Cell Science has deposited plasmids to generate fluorescently tagged proteins of key cellular ... stem cell (hiPSC) lines. The plasmids used to generate these lines have been deposited here with Addgene...With these plasmids researchers will be able to generate fluorescently tagged proteins of key cellular ...Microtubules 101786 ST6GAL1-mEGFP AICSDP-26 mEGFP Sialyltransferase 1 Golgi 109122 NPM1-mEGFP AICSDP-50 mEGFP ...164500 POLR2A-mEGFP AICSDP-117 mEGFP RPB1 RNA polymerase II 168798 TERF2-mEGFP AICSDP-77 mEGFP Telomeric...CTCF-binding sites 170818 PCNA-mEGFP AICSDP-116 mEGFP Proliferating cell nuclear antigen (PCNA) DNA replication ...Protocols The Allen Institute for Cell Science has several protocol videos available that may be helpful for...

... activator-like (TAL) effectors, scientists in several biological disciplines are coming closer to this...fingers recognize triplets and the FokI nuclease operates as a dimer, cutting in the spacer region between...triplet has a corresponding zinc finger, and interactions between zinc fingers within an array can reduce...zinc fingers, potential targetable sites in your average genome are every 500 bp. TAL effectors have some...history. Dr. Keith Joung’s lab at Massachusetts General Hospital also recently released a TALEN kit through... Zinc Finger Consortium with Dr. Voytas, has considerable experience in the DNA targeting field. A third...specific gene or your anxiously investigating gene therapy options for a rare disease in humans, TAL effectors...

...to LacZ, plus luciferase-2A-Cre recombinase 60226 : sgRNA cloning backbone with luciferase-2A-Cre recombinase...For cloning information, please view: Zhang Lab General Cloning Protocol 237.2 KB SpCas9 Plasmids The CRISPR...sgRNAs targeting KRAS , p53 , and LKB1 , plus luciferase-2A-Cre recombinase, and Kras G12D HDR template...of a single AAV vector ( AAV-KPL ) in the lung generated loss-of-function mutations in p53 and Lkb1 , as...plasmid contains two expression cassettes, Renilla luciferase-2A-Cre recombinase and sgRNAs targeting the mouse...plasmid contains two expression cassettes, Renilla luciferase-2A-Cre recombinase and an sgRNA targeted to LacZ...plasmid contains two expression cassettes, Renilla luciferase-2A-Cre recombinase and an sgRNA backbone for ...

...regions retaining molecular interactions and identify molecules interacting with those regions. Locus-specific...Lab web page . Figure 1: iChIP general scheme Figure 2: enChIP general scheme Fujii Lab Plasmids Individual...reveals genomic regulatory regions that physically interact with the Pax5 promoter in a chicken B cell line.../dsx023. PubMed . Identification of physical interactions between genomic regions by enChIP-Seq. Fujita...specific genomic regions retaining molecular interactions by the iChIP system using recombinant exogenous...

...Resources Overview of Viral Production Viruses are generated using standard methods that have been optimized...optimized for each specific virus in order to generate high quality preparations. After production, all virus...the University of Pennsylvania Vector Core. In general, titering is performed by the facility that produced...suggests that titers obtained using ddPCR are generally 2–3 fold higher than those achieved using the ...us . Full sequencing of the Viral Genome Next-generation sequencing is performed on viral genomes isolated...preparations are produced in 293T cells using the 2nd generation psPAX2 and pMD2.G packaging system developed ...dilutions of the lentiviral vector, harvesting cells several days later, and isolating genomic DNA. This DNA...

...Class I PIN1 Peptidylprolyl cis/trans isomerase, NIMA-interacting 1 PLXNB1 Plexin B1 PPP1CA Protein phosphatase...relationship. The content and map for this page were generated with the help of Dominic Esposito and the Frederick... related tyrosine kinase 3 FNT FNTA FNTB Farnesyltransferase, CAAX box FOS FBJ murine osteosarcoma viral...protein 2 ICMT Isoprenylcysteine carboxyl methyltransferase INSR Insulin receptor INSRR Insulin receptor-related... . Targeting RAS signalling pathways in cancer therapy. Downward J. Nat Rev Cancer. 2003 Jan;3(1):11-22...

...transduction of the trabecular meshwork following intracameral injection in mice, rats, and dogs. This serotype...serotype displays highly efficient transduction and lateral spread in the retina following subretinal injection...E531D) serotype has increased transduction and lateral spread in the retina following subretinal injection...Transduction optimization of AAV vectors for human gene therapy of glaucoma and their reversed cell entry characteristics...Display Exceptional Characteristics for Retinal Gene Therapy. Mol Ther . Jun 3;28(6):1464-1478. PMID: 32304666...

...and throughput over traditional tagging methods. Several different tagging techniques, as well as the plasmids...plasmids include Cre and FLP expression vectors, a general cloning plasmid, and a prebuilt Unc-32::GFP targeting...Auxin-inducible degron (AID) tagging with CRISPR/Cas to generate conditional alleles of essential nuclear and cytoplasmic...technique for use in Drosophila cells that uses PCR to generate both an expression cassette for the Cas9-programming...cells 3.3 MB Seydoux C. elegans Tagging System Geraldine Seydoux's lab has developed a systematic and scalable...elegans . This system uses a 10-day protocol, generates “clean” homozygous mutants with no co-integrated...

...18760 MSCV IRES Luciferase MSCV Plasmid for transgene expression; also expresses luciferase Lowe 9044 pMIG...gamma-retrovirus) is an RNA virus comprised of its genome and several structural and enzymatic proteins, including reverse...virus uses the reverse transcriptase in tow to generate a DNA provirus. This provirus then gets integrated...Sun 60683 pLXIN-Luc MoMSV Stable expression of luciferase in mammalian cells Wong Return to Top Do you ...

...produced in-house by Addgene scientists. They are generated using standard methods individually optimized ...optimized for each specific antibody in order to generate high quality preparations. Following production, all...using ImageJ software and a standard curve is generated. The final antibody concentration is calculated...labeled with the antibody and a species-specific horseradish peroxidase (HRP)-conjugated secondary antibody...passed stability testing, it may ship at room temperature. However, Addgene determines ideal shipping conditions...

...plasmids to test whether or not you've efficiently generated infectious virus, plasmids expressing both the...process for packaging these transfer vectors and generating infectious virus. Protocol: Lentivirus Transfer...coexpressed with tdTomato and Pac (puromycin N-acethyl-transferase) pCDH-CB-copGFP-T2A-iCre 72256 Expresses copGFP...72268 xpress gene of interest from the PGK (phosphoglycerate kinase I) promoter pCDH-CMV4 72284 Express... from pAdenoX from Clontech and can be used to generate adenovirus expressing a variety of reporters as...

...James M. Wilson AV-1-PV0105 105532-AAV1 pAAV.CMV.ffLuciferase.SV40 Control James M. Wilson AV-1-PV1917 105541...James M. Wilson AV-8-PV0105 105532-AAV8 pAAV.CMV.ffLuciferase.SV40 Control James M. Wilson AV-8-PV0146 105535...Karl Deisseroth AV-5-PV0105 105532-AAV5 pAAV.CMV.ffLuciferase.SV40 James M. Wilson AV-5-PV1090 105537-AAV5... M. Wilson AV-8-PV1302 105538-AAV8 pENN.AAV.TBG.PI.ffLuciferase.RBG James M. Wilson AV-8-PV3637 65015-...Karl Deisseroth AV-9-PV0105 105532-AAV9 pAAV.CMV.ffLuciferase.SV40 James M. Wilson AV-9-PV0109 105533-AAV9...Karl Deisseroth AV-2-PV0105 105532-AAV2 pAAV.CMV.ffLuciferase.SV40 James M. Wilson AV-1-PV3365 105554-AAV1...

...maintains a Standard Operating Procedure (Link opens in a new window) with general principles for working... in well-defined cellular sub-populations has generally been limited to single-component cellular definitions... cotransmission evokes a delayed excitation in lateral dorsal striatal cholinergic interneurons. Elife... The central amygdala controls learning in the lateral amygdala. Nat Neurosci. 20(12):1680-1685. PubMed...

...proteins then express and process the CRISPR loci to generate CRISPR RNAs (crRNAs). Through sequence homology...or repression. Combined with the power of next-generation sequencing, CRISPR is a robust system for genome-wide...RNA. Inosine is translated as guanosine, so the overall effect is an A->G change. Since this method targets...technologies. Biotechnology companies are exploring therapeutic applications of CRISPR to treat genetic disease...ethics associated with CRISPR in agricultural and therapeutic applications. Despite the ethical controversies...

...expectations communicated by email will supersede the general guarantees above. We will have our expert viral...Production of a Packaged on Request material is generally ready for shipment within 8 weeks after the approval...queue is too high and we do not expect to meet the general turnaround time, Addgene will notify you by email...expectations communicated by email will supersede the general estimated turnaround time above. After starting...