...serotype is demonstrated to have enhanced transduction of the retina following intravitreal injection in...also displays enhanced transduction of the trabecular meshwork following intracameral injection in mice, ...enhanced transduction of the retina following subretinal administration and enhanced transduction and spread...) serotype shows enhanced transduction of the retina following intravitreal injection in mice and macaque...macaque. This serotype also demonstrates enhanced transduction of cells in culture for ex-vivo conditioning...Anterior Chamber Results in Efficient Transduction of Trabecular Meshwork in Mouse and Rat. PLoS One ....High-Efficiency Transduction of Primary Human Hematopoietic Stem/Progenitor Cells by AAV6 Vectors: Strategies for ...

...mutation, the best HR template for transfection is probably a single stranded DNA (ssDNA) oligo which usually...discussing conversion tract length in drosophila. This 1998 study suggests mammalian cells' tract lengths seem ...positively transfected. Alternatively, you can use a selection marker to select transfected cells (for...recommends using the Epicentre QuickExtract solution to perform this step to extract genomic DNA from cells. ...50ul PCR reaction. The gDNA extracted in this way is usually more concentrated than using other kits like...nickase doesn't have the ability to induce double-stranded breaks like the wildtype Cas9 does. There are ... The human U6 promoter prefers a 'G' at the transcription start site to have high expression, so adding...

...Plasmids 101: E. coli strains for protein expression Plasmids 101: Common Lab E. coli Strains Why Bacteria? E...pdCas9-bacteria 44249 pTetO Anhydrotetracycline Stanley Qi Anhydrotetracycline inducible expression of catalytically...light. pCph8 50552 pLtetO-1 Anhydrotetracycline Jeffrey Tabor Anhydrotetracycline inducible expression of ...testing transcription factor binding to reporter plasmid pH3U3-zif268 (omega) . Transcription factors ...Keith Joung Transcription factor testing: Kit contains five plasmids and one bacterial strain for performing... answer many interesting questions. Does my transcription factor bind to a particular sequence? Do these...pwtCas9-bacteria 44250 CRISPR Stanley Qi Anhydrotetracycline inducible expression of wild-type Cas9 from...

...adenovirus terms Adenoviral Strains Bacterial strains for generating adenovirus. ID Strain Description PI 16398...containing transgene under CMV promoter Vogelstein 16404 pAdTrack Shuttle For production of GFP-trackable viruses...Zhang 50957 RedTrackCMV Shuttle For production of mRFP-trackable viruses containing transgene under CMV ...genome. Disadvantages of adenoviruses include transient transgene expression, as well as the substantial immune...viruses containing transgene under a chosen promoter Vogelstein 16405 pAdTrack-CMV Shuttle For production...production of GFP-trackable viruses containing transgene under CMV promoter Vogelstein 16407 pAdEasy 2-GFP beta-gal...Collection of adenovirus shuttle vectors and strains Viral...

...expression, multicistronic design enables tracking of viral transduction. Also see plasmid 46948 . Elledge 44011...knockdown, multicistronic design enables tracking of viral transduction. See article for more similar plasmids...Addgene lentiviral packaging, envelope, and transfer plasmids Viral...Libraries All Viral Plasmids Packaging Plasmids Transfer Plasmids Lentiviruses, a subset of retroviruses...types of viruses used in research. Lentivirus can transduce both dividing and non-dividing cells without a...stably into the host genome, enabling long term transgene expression. There are some safety considerations...encodes the viral genome (typically called the transfer plasmid ). This table lists common lentiviral ...

...pOpen-EcoRIM M.EcoRI DNA methyltransferase 165517 pOpen-Eco31IA Eco31IA DNA methyltransferase 165518 pOpen-NotIM...pOpen-NotIM M.NotI DNA methyltransferase 165527 pOpen-Eco31IB Eco31IB DNA methyltransferase 165536 pOpen-PstIM...pOpen-PstIM M.PstI DNA methyltransferase 165538 pOpen-SfiIM M.SfiI 165538 DNA methyltransferase 165547 pOpen-SapIM1...pOpen-SapIM1 M1.SapI DNA methyltransferase 165557 pOpen-SapIM2 M2.SapI DNA methyltransferase 165558 pOpen-EcoRVM...pOpen-EcoRVM M.EcoRV DNA methyltransferase 165577 pOpen-XbaIM M.XbaI DNA methyltransferase 165578 pOpen-NcoIM ...32 single-stranded DNA-binding protein 165549 pOpen-ETSSB Extreme Thermostable Single-Stranded DNA-Binding...Polymerases Restriction Endonucleases Reverse Transcriptases RNA Polymerases Other Enzymes (Link opens in...

...Adenoviral High transduction efficiency, but do NOT integrate into host genome pAdTrack-CMV - Shuttle...production AdEasier®-1 cells (strain) - Bacterial strain that contains AdEasy®-...miniTurbo in the mammalian nucleus pSF3-ultraID - Expresses ultraID in mammalian cells with the Tet-On or...expression Find yeast transformation plasmids with G418 selection Find nematode transgenesis plasmids with Neomycin...Backbones Mammalian CMV, SV40, EF1a, CAG, Ubc Transient expression - Gradia Lab plasmids for mammalian...Find more Tet-inducible empty backbones on our Tetracycline (Tet) Inducible Expression page Bacteria Lac...gene with N-terminal GST tag and TEV cleavage Transient expression - Gradia Lab plasmids for bacterial...

...protein translation machinery. To expand the genetic code, 4 major changes to the standard translation machinery... Mehl 85496 pDule-Tet2.0 Tetrazine2.0 tRNA synthetase M. jannaschii Tetrazine 2.0 Bacterial TAG Ryan Mehl...Mehl 85497 pDule2-Tet2.0 Tetrazine2.0 tRNA synthetas M. jannaschii Tetrazine 2.0 Bacterial TAG Ryan Mehl...barkeri Tetrazine 3.0 Bacterial TAG Ryan Mehl 174081 pAcBac1-NES-Flag-R2-84-MbRS Tetrazine3.0 NES-Flag-R2... TAG Edward Lemke Browse Strains The table below highlights bacteria strains that have been modified to...Addgene's guide to plasmids and strains useful for expanding the genetic code in bacteria and mammalian...Rinehart Lab reagents tRNA/Synthetase Pairs Bacterial Strains Codon Modified Constructs The genetic code for ...

...special trans-activating crRNA (tracrRNA) with homology to the short palindromic repeat. The tracrRNA helps... double stranded DNA target and cleaves both strands near the PAM. (6) The crRNA:tracrRNA:Cas9 complex...: dCas9 fused to transcriptional activators or repressors can modulate transcription in a reversible manner...have long utilized CRISPR as a powerful defensive strategy against viral invaders, and this system is proving...palindromic repeat sequences. (1) The CRISPR array is transcribed to make the pre-CRISPR RNA (pre-crRNA). (2) The...together separate the individual crRNAs. (3) The tracrRNA and Cas9 nuclease form a complex with each individual...individual, unique crRNA. (4) Each crRNA:tracrRNA:Cas9 complex seeks out the DNA sequence complementary...

...so cells must be lysed or the substrate must be delivered intracellularly in order to measure luminescence...Bioluminescence Resonance Energy Transfer (BRET) CalFlux VNT : An intracellular calcium sensor created by fusing...including transcriptional reporters for NF-kb, TGF-b, c-Myc, p53 and MAPK/JNK transcriptional reporters...collection of luciferase plasmids for assays of transcriptional regulation and bioluminescent reporters. Plasmid...insects. Luciferase catalyzes the oxidation of a substrate, leading to the emission of photons. The robust... protein half-lives, making them useful as transcriptional reporters. However, they are not secreted, ...regulatory elements, such as promoters, enhancers and untranslated regions, or the effect of mutations of these...

...reprogramming factors Efficient germ-line transmission obtained with transgene-free induced pluripotent stem cells...Reprogramming Plasmids Differentiation and Transdifferentiation Plasmids Stem Cell Research Plasmids Stem...methods with the goal of helping readers choose a strategy suited to their research. Once you know which ...differentiated somatic cell into another via transdifferentiation. This method of cell lineage conversion ...technology. Browse Addgene's collection of Transdifferentiation Plasmids below. Many of the techniques for... for reprogramming, differentiation, and transdifferentiation utilize plasmids to increase expression ...human Klf4, Oct4, c-Myc, and Sox2 as VP-16 transcriptional activating fusions in single lentiviral vectors...

...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type... opposite strands of your DNA in close proximity. These double nicks create a double-strand break (DSB...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ... BspQI Transfection Puromycin yes, cut Ji-Long Liu 45946 pU6-BbsI-chiRNA dU6 BbsI Transfection none, need...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...functional CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...non-homologous end joining (NHEJ). Double nicking strategies reduce unwanted off-target effects. Nickase mutants...

...Plant transcription factors for Agrobacterium-mediated transient expression or plant transformation - Silin...parts for assembling plant transformation vectors - Daniel Voytas. Transcription Factors The following collections...expression in plants: Maize Transcription Factor ORFeome - John Gray. Rice Transcription Factor ORFeome - John...Plant Base Editors Save Time with Transient Plant Leaf Transformations RNA Interference in Plant Biology...Figshare - Plant Illustrations (Link opens in a new window) - Collection of illustrations and pictures of... Addgene Blog: Plant Biology CRISPR Cloning Transcription Factors RNAi Empty Backbones Fluorescent Proteins...Engineered biocontainable RNA virus vectors for non-transgenic genome editing across crop species and genotypes...

...sites are on separate DNA molecules (in trans), a translocation event is generated between the target sites...85577 pTC-CMV-Tet CreER expression and tetracyclin-dependent transgene/shRNA expression ApoE.HCR.hAAT Mammalian... pTC-ApoE-Tet CreER expression and tetracyclin-dependent transgene/shRNA expression ApoE.HCR.hAAT Mammalian...plasmids can be useful for lineage tracing, evaluating infection/transfection efficiency, selecting desired...of rearrangements: inversion, deletion, and translocation. Figure 1. Recombination outcomes are determined... If the target sites (loxP) are on the same DNA strand in opposite directions, recombination results in...reversed. B) If the target sites are on the same DNA strand (in cis) in the same direction, the sequence between...

...For easy-to-transfect cell types (e.g. HEK293 cells), transfection with standard transfection reagents may...mammalian cell line that can be transfected at high efficiency Lentiviral transduction Cas9 and gRNA can be present...present in a single lentiviral transfer vector or separate transfer vectors May contain reporter gene ...screens using CRISPR AAV transduction CRISPR elements are inserted into an AAV transfer vector and used to ...ribonucleoprotein (RNP) complexes are advantageous due to the transient Cas9 expression. The table below summarizes the...details, see Plasmids 101: Mammalian Vectors ) Transient or stable expression of Cas9 and/or gRNA in a ... mammalian cell lines Useful for difficult to transfect cell types, can also be used in vivo A common ...

... recoded strain of E. coli ( 68306 ) with the optimized phosphoserine orthogonal translation system ( ...Plasmid 69118 E17TAG GFP zeo resistance Strain 34929 BL21ΔserB Strain 52055 EcAR7 Plasmid 112031 pNAS1b split...phosphorylation is one of the most abundant forms of posttranslational modifications in cells and research into its..., Science 2013) and improving the orthogonal translation system to make singly or multiply phosphorylated...pSerOTS. Recently the Rinehart lab published a new strain of recoded E.coli (rEcoli XpS 192872 ) and an optimized... optimized phosphoserine orthogonal translation system (pSerOTS-C1* (V70) 188537 ). These reagents are...phosphorylation sites (“phosphosites”) consist of a central phosphoserine site flanked by 15 amino acids on...

...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type... opposite strands of your DNA in close proximity. These double nicks create a double-strand break (DSB...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ...functional CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...base editors convert C->T (or G->A on the opposite strand) within a small editing window specified by the...guanosine to create A->G (or T->C on the opposite strand) mutations. ID Plasmid Promoter PI Publication ...

...Tools Tetracycline Transactivators Affinity Purification Neurophysiology Cell Ablation Tetracycline Transactivators...These AAV encode tetracycline-inducible tools/controls and tetracycline transactivators that can be used...service encoding tet-off transactivators and tools for affinity purification (TRAP). Viral...used with tetracycline (tet)-inducible expression systems . ID Name Expression System Activity Serotype...CAG-driven, constitutive Expression of the tet-off transactivator (tTA) 2 Gradinaru 99120 pAAV-ihSyn1-tTA Inducible... promoter (ihSyn) Expression of the tet-off transactivator (tTA) with a positive feedback loop for amplified...ihSyn) Cre-dependent expression of the tet-off transactivator (tTA) with a positive feedback loop for amplified...

...- Tatjana Sauka-Spengler Lab Lineage Tracing Cell lineage tracing in multicellular systems typically relies...Lewis Lab PhOTO Zebrafish: A Transgenic Resource for In Vivo Lineage Tracing during Development and Regeneration...Periklis Pantazis Lab Multibow: digital spectral barcodes for cell tracing - Sean Megason Lab Zebrabow - Albert... (~3 months), ease of care and breeding, and transparency of the embryo and larvae, it is easy to culture...standard toolbox of genetic methods, such as Tol2 transposons, Cre-mediated site-specific recombination, morpholinos...emerged as a model system for human disease and translational research, including the study of cancer, inflammation...-Cas9 system - Shuo Lin Lab Re-purposed Ac/Ds transgenic system enabling CRISPR/dCas9-interference (CRISPRi...

...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type... opposite strands of your DNA in close proximity. These double nicks create a double-strand break (DSB...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ...functional CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...Base editors convert C->T (or G->A on the opposite strand) within a small editing window specified by the...guanosine to create A->G (or T->C on the opposite strand) mutations. CRISPR-X, a base-editing system from...