...Flpo recombinase Recombinases Zhang 121675 pAAV-EF1a-fDIO-Cre EF1a Flp-dependent Cre recombinase Recombinases...EYFP Control Gradinaru 105547 pENN.AAV.EF1a.eGFP.WPRE.rBG EF1a EGFP Control Wilson Recombinases 24593 AAV-pgk-Cre...expression Recombinases Larsen 105553 pENN.AAV.hSyn.Cre.WPRE.hGH Syn Cre expression Recombinases Wilson 107788...mCherry expression Recombinases Deisseroth 55636 pAAV-EF1a-Cre EF1a Cre expression Recombinases Deisseroth 55637... expression Recombinases Deisseroth 55638 pAAV-EF1a-vCre EF1a VCre expression Recombinases Deisseroth ...expression Recombinases Zeng 51669 AAV phSyn1(S)-FlpO-bGHpA Syn Flpo expression Recombinases Zeng 183412...possible) in vivo testing, and full sequencing of the final viral vector preparation. In some cases, in vivo...

...viruses (AAV) are small viruses originally discovered as contaminants of adenovirus stocks. One major ...amplification system Gradinaru 129024 pProvector_AAV2_586_PLGLAR AAV2_586_PLGLAR Recombinant AAV2 rep/cap plasmid...low immune response and low toxicity. Although recombinant AAV does not integrate into the host genome, ...Cap controlled by tTA-TRE amplification system Gradinaru 103006 pUCmini-iCAP-PHP.S PHP.S AAV packaging ...Cap controlled by tTA-TRE amplification system Gradinaru 127847 pUCmini-iCAP-PHP.V1 PHP.V1 AAV packaging...Cap controlled by tTA-TRE amplification system Gradinaru 175004 pUCmini-iCAP-AAV.CAP-B10 B10 AAV packaging...Cap controlled by tTA-TRE amplification system Gradinaru 175005 pUCmini-iCAP-AAV.CAP-B22 B22 AAV packaging...

...displays enhanced transduction of the retina following subretinal administration and enhanced transduction...transduction and lateral spread in the retina following subretinal injection. It was identified by John ...transduction and lateral spread in the retina following subretinal injection as compared to the parental...Heparan Sulfate Binding on Transduction of Retina by Recombinant Adeno-Associated Virus Vectors. J Virol ...may also like: Viral Service: All AAV Viviana Gradinaru PHP AAV Serotypes Blog: Viral Vectors As part ...possible) in vivo testing, and full sequencing of the final viral vector preparation. These viral vector preparations...delivered directly to the brain parenchyma or subretinal space Serotypes The following serotypes were ...

...sequence in the 3' UTR Carl Novina 33154 pAC-Luc Firefly Creation of N-terminal Firefly fusions in Drosopholia...luciferase reporter. These plasmids improve over the original STARR-seq plasmids by using the bacterial ORI ...which was a source of false positives with the original STARR-seq method. Learn more about STARR-seq in...87067 pcDNA3.1-ccdB-Nanoluc NanoLuc® Creation of N-terminal Nanoluc fusions using Gateway cloning Mikko Taipale... pLenti6.2-ccdB-Nanoluc NanoLuc® Creation of C-terminal Nanoluc fusions using Gateway cloning. Lentival... pLenti6.2-Nanoluc-ccdB NanoLuc® Creation of N-terminal Nanoluc fusions using Gateway cloning. Lentiviral...87070 pcDNA3.1-Nanoluc-ccdB NanoLuc® Creation of N-terminal Nanoluc fusions using Gateway cloning Mikko Taipale...

...luciferase-2A-Cre recombinase 60226 : sgRNA cloning backbone with luciferase-2A-Cre recombinase Vectors for ... to NeuN, plus Cre recombinase 60228 : sgRNA targeted to LacZ, plus Cre recombinase 60229 : sgRNA cloning... backbone with Cre recombinase 60230 : sgRNA targeted to NeuN, plus Cre recombinase-2A-EGFP-KASH 60231...plasmids. CRISPR loci in microbial hosts contain a combination of CRISPR-associated (Cas) genes as well as non-coding...consists of three components: A nucleolytically inactive Cas9-VP64 fusion A sgRNA incorporating two MS2...targeting efficiency. These plasmids use the inverted terminal repeats (ITR) from AAV serotype 2. SaCas9 only...KRAS , p53 , and LKB1 , plus luciferase-2A-Cre recombinase, and Kras G12D HDR template 60225 : control for...

...pCAG-T7-TALEN(Sangamo)-Destination Pawel Pelczar pCAG-T7-TALEN(Sangamo)-Destination constructs were designed...TALEN(Sangamo)-FokI-KKR-Destination 40132 pCAG-T7-TALEN(Sangamo)-FokI-ELD-Destination Golden Gate TALEN 2.0...new destination vectors can be used to create TALE-transcription factors and are used at the final assembly...pC-GoldyTALEN and RCIsricpt-GoldyTALEN are designed as destination vectors for the Voytas lab Golden Gate TALEN ...-BB and pTAL6-BB are designed as alternative destination vectors for the Golden Gate TALEN kit (Bogdanove...expression vector resulting in shorter N- and C-terminal tal protein segments (136AA and 63AA, respectfully...several later studies. pCAG-T7-TALEN(Sangamo)-Destination vectors are available with homodimeric or enhanced...

...pAAV-mDlx-NLS-mRuby2 Dlx mRuby2 Control Gradinaru 104055 pAAV-CAG-eYFP CAG EYFP Control Gradinaru 104061 CAG-NLS-GFP CAG...light-inducible recombinase iCreV Zeng 140136 pAAV-EF1a-iDreV EF1a light-inducible recombinase iDreV Zeng ... Engineered systemic constructs from Viviana Gradinaru's lab at Caltech. Viral...engineered systemic serotypes from the Viviana Gradinaru laboratory and the Caltech CLOVER Center . As ...possible) in vivo testing, and full sequencing of the final viral vector preparation. Systemic Delivery AAV ...NHP, non-human primate. Image courtesy of the Gradinaru Lab and the Caltech CLOVER Center and was adapted...CAG NLS-GFP Control Gradinaru 105530 pAAV.CMV.PI.EGFP.WPRE.bGH CMV EGFP Control Wilson 105547 pENN.AAV....

...nickase or catalytically inactive “dead” Cas9 (dCas9) to a cytidine deaminase like APOBEC. Base editors...target sequence present in the gRNA. CRISPR was originally employed to knock out target genes in various... gRNA binding that shifts the molecule from an inactive, non-DNA binding conformation into an active DNA-binding... that increase Cas9 proofreading and target discrimination. All three high fidelity enzymes generate less...resulting population of cells will contain some combination of wild type alleles, NHEJ-repaired alleles, ...creating an A to G (or T to C) change. Adenine DNA deaminases do not exist in nature, but have been created... of the Escherichia coli TadA, a tRNA adenine deaminase. Like cytosine base editors, the evolved TadA...

...the N-terminal portion of split mCherry, while a phosphobinding domain is fused to the C-terminal mCherry... of kinases of interest by mass spectrometry in a technique called serine-oriented library/kinase-library...overall improved ease of use when expressing recombinant phosphoproteins. These attributes make rEcoli...111705-8 ) is available from Addgene. Screening kinases. The mode #1 phosphosite library can also be synthesized...enhanced biological tolerance. Mohler K, Moen JM, Rogulina S, Rinehart J. Mol Syst Biol 2023 Aug 8;19(8):...interactions. Barber KW, Muir P, Szeligowski RV, Rogulina S, Gerstein M, Sampson JR, Isaacs FJ, Rinehart... Nat Biotechnol. 2018 Aug;36(7):638-644. PubMed Kinase Substrate Profiling Using a Proteome-wide Serine-Oriented...

...absence of unexpected DNA contaminants. The data is also used to determine recombination rates in FLEX/DIO/fDIO...Endotoxin contamination in vector preparations can alter the immunogenic properties of the final product...mycoplasma contamination. To date, Addgene has never had a case of mycoplasma contamination. In the event...is performed on viral genomes isolated from the final AAV preparation. Sequencing results are analyzed...particularly in large animal studies. Endotoxin contamination is minimized by using an endotoxin-free plasmid.... To minimize the immunogenic properties of the final vector preparation, the quantity of gram-negative...Takara, and is routinely tested for mycoplasma contamination using mycoplasma detection kits. The cell line...

...Verdoes Backbones for mouse recombinant antibodies A panel of recombinant monoclonal antibodies against... Collection Addgene distributes ready-to-use recombinant antibodies made from plasmids in our collection...alternative approach is to use synthetically produced recombinant antibodies, created by cloning antibody components...Some scientists choose to work with full length recombinant monoclonal antibodies (R-mAbs) expressed in mammalian...research applications from the NeuroMab/Trimmer Lab Recombinant mAb Collection . Once the plasmids are transiently...of production due to their small size. These recombinant antibody fragments can be used in experiments...tool kit for rapid cloning and expression of recombinant antibodies. Sci Rep. 2014 Jul 30;4:5885. Andrew...

...growth and proliferation. mTOR, a serine/threonine kinase, is found in two complexes, mTORC1 and mTORC2... and multiple familial cancers are caused by inactivating mutations in negative regulators of mTORC1. ...homolog LKB1 Also known as STK11; serine/threonine kinase 11 mTOR Mechanistic target of rapamycin NF1 Neurofibromin...S6K Also known as RPS6KB1; Ribosomal protein S6 kinase B1 TSC1/2 TSC1 TSC2 Tuberous sclerosis VHL Von ...LST8 homolog GSK3 GSK3A GSK3B Glycogen synthase kinase 3 MDM2 MDM2 proto-oncogene mSin1 Also known as ...as MAPKAP1; Mitogen-activated protein kinase associated protein 1 mTOR Mechanistic target of rapamycin ...; tumor protein p53 PDK1 Pyruvate dehydrogenase kinase, isozyme 1 PI3K Catalytic Subunits PIK3CA PIK3CB...

...Antibodies produced this way are called recombinant antibodies. Recombinant antibodies are created by cloning... time and labor intensive. Recombinant expression validation A recombinant expression strategy is used...cultured immune cells, or through the use of recombinant plasmids. Animals - Antibodies produced in animals... of a single antibody with little variation. Recombinant plasmids - Antibodies can be produced from plasmids... and can either be used while suspended in the original media or isolated and placed in a buffer. Antibody... binding site. This process, known as V(D)J recombination, allows for a wide variety of antibodies to ...abundant isotype in blood, also found in tissues; predominant isotype in secondary immune response Monomer ...

...3rd 6-25 11,108 Borodina Yeast Transporter Libraries 153101-153106 Knockout Yeast Borodina N/A 1 Varies ...Human Ebert 3rd 4 2,852 Bonifacino Lab Human ubiquitination-related proteins CRISPR KO library 174592 Knockout...contained in the library represent all 4096 (4 6 ) combinations of 6 nucleotides. Chong Zhang E. coli Genome-wide... Human Weissman 3rd 6 Varies Enriched subpools (kinase, nuclear, ribosomal, cell cycle) 51043 — 51048 ... 84,963 Human CRISPR Knockout Pooled Library (Gattinara) 136986 Knockout Human Doench 3rd 2 40,964 Ingolia...Library 140633 Knockout Human Wei 3rd 3 58,630 Human Kinase Domain-Focused CRISPR Knockout Library 117725 Knockout...

...to brainstorm practical strategies for using combinations of AAV-based molecular tools to achieve targeted...Cambridge, MA, 02139 Organizing Committee Chair: Dr. Melina Fan, Co-Founder and CSO, Addgene Organizing Committee...Dr. Constance Cepko, Harvard Medical School Dr. Melina Fan, Dr. Susanna Bachle, Dr. Karen Guerin, Dr. ...Research Campus Kenton Woodard Penn Vector Core Melina Fan, Karen Guerin, and Leila Haery from Addgene...Friday, September 21 9:00 AM Breakfast and Welcome Melina Fan 9:30 AM Cre and Flpo Katherine Matho 10:15 ...Regulation Ben Deverman 2:00 PM Cre-DOG and Combinatorial Strategies Connie Cepko 2:45 PM Break 3:00 PM...Field Kenton Woodard 4:00 PM Wrap Up & Next Steps Melina Fan/Leila Haery 5:00 PM Adjourn Meeting Preparation...

...vector for insertion of N-terminal domain (SgrAI or AgeI with ClaI) and C-terminal domain (FseI PacI). Grow...modules are orthogonal, researchers can use any combinations of modules as is, allowing plug-and-play application...

...backbone-based Gateway destination vectors (pGCS), bearing either amino- or carboxyl-terminal tags, including...zebrafish research. China Zebrafish Resource Center (CZRC) - A resource center based in China focused on collecting...Tol2 transposons, Cre-mediated site-specific recombination, morpholinos, zinc finger nucleases, TALENS,...vectors for cell-type specific conditional gene inactivation - Jeffrey Essner Lab GeneWeld Vectors for Targeted...origin in zebrafish can be found below: Optimized QF-binary expression system for use in zebrafish - Bret Pearson...zebrafish Whole organism lineage tracing by combinatorial and cumulative genome editing - Jay Shendure...toolkit - Tatjana Sauka-Spengler Lab. A genetic binary system for cell- and compartment-specific biotin...

...between mouse and human Jonathan Ting Viviana Gradinaru 163509-PHPeB CN1839-rAAV-hSyn1-SYFP2-10aa-H2B-...scRNA-seq applications. Jonathan Ting Viviana Gradinaru 164450-PHPeB CN1851-rAAV-hI56i-minBglobin-iCre...forebrain GABAergic interneurons. For use in combination with Cre-dependent transgenic mouse lines or ...dependent viral vectors. Jonathan Ting Viviana Gradinaru 37825-CAP-B10 pAAV-CAG-GFP AAV expression of GFP...GFP from the CAG promoter Edward Boyden Viviana Gradinaru 37825-CAP-B22 pAAV-CAG-GFP AAV expression of GFP...GFP from the CAG promoter Edward Boyden Viviana Gradinaru 37825-CAP-MaCPNS1 pAAV-CAG-GFP AAV expression ...GFP from the CAG promoter Edward Boyden Viviana Gradinaru 37825-CAP-MaCPNS2 pAAV-CAG-GFP AAV expression ...

...CDK6 Cyclin-dependent kinase 4 or 6 CHK1 Checkpoint kinase 1 CHK2 Checkpoint kinase 2 Cop-1 Ring finger ...factor 1 ATM ATM serine/threonine kinase ATR ATR serine/threonine kinase B99 G-2 and S-phase expressed 1...PMAIP1 p14ARF Cyclin-dependent kinase inhibitor 2A p21 Cyclin-dependent kinase inhibitor 1A p48 Damage-specific...to activate transcription, and they also have a dominant negative effect on functional p53 through oligomerization...

...Homologous Recombination (HR)? How long should the homology arms be for efficient recombination? The most...Dr. Le Cong, the first author of a seminal CRISPR article from Dr. Feng Zhang's Lab, has assembled a ...Cong Dr. Le Cong, the first author of one of the seminal articles from Dr. Feng Zhang's Lab ( Science , ...to learn more. CRISPR Design FAQs Homologous Recombination (HR) FAQs At the Bench FAQs CRISPR Design FAQs... to express the Cas9n (nickase). Homologous Recombination (HR) FAQs Can you summarize HR template vector...specific mutations or insertions by Homologous Recombination (HR), what is the length of each homology arm...destroy the BbsI sites, a double digest should discriminate between positive and negative clones. Clones...