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  1. CRISPR References and Information

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    ...and references, including gRNA design tools, CRISPR FAQs, and community forums.... CRISPR References and ...and Information CRISPR References and Information Addgene Resources CRISPR Software General Resources CRISPR...kits. CRISPR Software Sanger Indel Analysis ICE (Inference of CRISPR Edits) This open access tool from Synthego...automate and optimize gRNA and HDR donor design. The current version supports SpCas9, SpCas9-VQR, and enAsCas12a...opens in a new window) This tool helps design (10 different prediction scores), clone (primer design), and...inactivation, NHEJ, and HDR. Includes a wide range of reference genomes, including animals, plants, bacteria, ...
  2. Luciferase Plasmid Collection

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    ...74444 pLenti.PGK.blast-Renilla_Luciferase Renilla PGK Lentiviral expression of Renilla luciferase Reuben Shaw...Root 118016 pLX313-Renilla luciferase Renilla EF1α Lentiviral expression of Renilla luciferase William...pAAV-CAG-RLuc Renilla CAG AAV expression of Renilla luciferase Mark Kay 83280 pscAAV-CAG-RLuc Renilla CAG Vector...Orzaez 68215 pEGB 35S:Renilla:Tnos (GB0109) Renilla 35S Transcriptional unit for Renilla luciferase expression...depending on the application, and different luciferases require different substrates and detection wavelengths... than Renilla luciferase, but is more susceptible to enzyme inhibition. Both firefly and Renilla luciferase...12179 pIS1 Renilla Insertion of target sequence in the 3' UTR David Bartel 12177 pIS2 Renilla Insertion...
  3. Biosensor AAV Preps

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    ...Constitutive 1 Loren Looger 137955 pAAV.CAG.iAChSnFR CAG iAChSnFR none Constitutive 1, 9 Loren Looger 121922..., rg* Loren Looger 176753 AAV-mDlx-jGCaMP8f-WPRE Dlx jGCaMP8f none Constitutive 1, 9, rg* Loren Looger...Constitutive 1, 9 Loren Looger 179254 AAV-CAG-jGCaMP8f-WPRE CAG jGCaMP8f none Constitutive 1, 9, rg* Loren Looger..., rg* Loren Looger 176755 AAV-mDlx-jGCaMP8s-WPRE Dlx jGCaMP8s none Constitutive 1, 9, rg* Loren Looger...Constitutive 1, 9 Loren Looger 179256 AAV-CAG-jGCaMP8s-WPRE CAG jGCaMP8s none Constitutive 1, 9, rg* Loren Looger..., rg* Loren Looger 176754 AAV-mDlx-jGCaMP8m-WPRE Dlx jGCaMP8m none Constitutive 1, 9, rg* Loren Looger... 5, 9 Loren Looger 176757 AAV-EF1a-jGCaMP8m-WPRE EF1a jGCaMP8m none Constitutive 1, 9, rg* Loren Looger...
  4. Plasmids for Stem Cell Research

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    ...Research Reprogramming Plasmids Differentiation and Transdifferentiation Plasmids Stem Cell Research Plasmids...unspecialized cells with the capacity to differentiate into many different cell types in the body during development...can be directly differentiated into specific somatic stem cells or fully differentiated cell types. Browse...collection of Differentiation Plasmids below. Alternatively, it is also possible to directly differentiate one ...one differentiated somatic cell into another via transdifferentiation. This method of cell lineage conversion... of Transdifferentiation Plasmids below. Many of the techniques for reprogramming, differentiation, and... , SOX2 , MYC , KLF4 , LIN28 . Differentiation and Transdifferentiation Plasmids Browse the table below...
  5. Plan Your Experiment

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    ...many different Cas proteins available, and researchers can adapt these experiments for different organisms...experiment. Created with BioRender.com. Select Your Desired Genetic Manipulation Different genetic manipulations...Activation/interference Needs good primer design Western blot Knockouts Activation/interference Needs a ...Expression and Delivery Validate Genetic Edit References CRISPR ( C lustered R egularly I nterspaced S...have expanded the CRISPR toolbox to include many different types of edits, including point mutations, base...manipulations require different CRISPR components. Selecting the genetic manipulation you plan to perform will...Base editor (dCas9 or Cas9 nickase fusions with different base editing proteins) Many base editors fused...
  6. Penn Vector Core Partnership with Addgene

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    ...Biosensor Loren Looger AV-1-PV2724 98931-AAV1 pAAV.hSyn.Flex.iGluSnFr.WPRE.SV40 Biosensor Loren Looger AV...Biosensor Loren Looger AV-5-PV2724 98931-AAV5 pAAV.hSyn.Flex.iGluSnFr.WPRE.SV40 Biosensor Loren Looger AV...Biosensor Loren Looger AV-9-PV2724 98931-AAV9 pAAV.hSyn.Flex.iGluSnFr.WPRE.SV40 Biosensor Loren Looger AV...Control Loren Looger AV-1-PV3661 98927-AAV1 pENN.AAV.CAG.Flex.GFPsm_myc.WPRE.SV40 Control Loren Looger ....2A.nlsmCherry.loxP.WPRE.SV40 Lawrence Cohen AV-1-PV3509 98925-AAV1 pAAV.CAG.Ruby2sm-Flag.WPRE.SV40 Loren Looger AV-1-PV3854... list of Addgene's viral vectors See Addgene's Current AAV inventory , which includes all viral vectors...AAV1 pAAV.CAG.Flex.iGluSnFr.WPRE.SV40 Biosensor Loren Looger AV-1-PV2816 100835-AAV1 pAAV.CAG.Flex.GCaMP6f.WPRE.SV40...
  7. Antibody Guide

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    ...heavy chain constant region” or Fc. Different isotypes play different roles in the immune response, and ...of colors activated by different wavelengths, allowing for easy differentiation between multiple antibodies...roughly fifty different fluorophore colors in flow cytometry, but the largest panels used currently are in the...Antibodies are typically produced in one of three different methods: in animals, in cultured immune cells,...contain two Fab fragments, each recognizing a different epitope, bound by short peptide linkers. Diabodies...used as an antimicrobial agent. Other chemical interferences with conjugating reactions are possible. It ...chemiluminescent reaction in the presence of several different substrates. The luminescence produced by the reaction...
  8. Retrograde AAV viral preps

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    ... CAG GFP Control Loren Looger 116870 pAAV-CAG-H2B tdTomato CAG tdTomato Control Loren Looger 24593 AAV-pgk-Cre...Calcium sensor Loren Looger 176749 AAV-Syn-H2B-jGCaMP8s-WPRE Syn H2B-GCaMP8s Calcium sensor Loren Looger 176750...Calcium sensor Loren Looger 176751 AAV-CamKIIa-jGCaMP8m-WPRE CamKII GCaMP8m Calcium sensor Loren Looger 176752...Calcium sensor Loren Looger 176753 AAV-mDlx-jGCaMP8f-WPRE Dlx GCaMP8f Calcium sensor Loren Looger 176755...Calcium sensor Loren Looger 176756 AAV-EF1a-jGCaMP8f-WPRE Ef1a GCaMP8f Calcium sensor Loren Looger 176757...Calcium sensor Loren Looger 176758 AAV-EF1a-jGCaMP8s-WPRE EF1a GCaMP8s Calcium sensor Loren Looger 179254...Calcium sensor Loren Looger 179255 AAV-CAG-jGCaMP8m-WPRE CAG GCaMP8m Calcium sensor Loren Looger 179256...
  9. CRISPR Guide

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    ...created using BioRender.com (Link opens in a new window) . Addgene Website & Blog References Website CRISPR..., A., Herreno-Pachón, A., Benincore-Flórez, E., Karunathilaka, A., & Tomatsu, S. (2024). Current strategies...Alternatives Resources Plan Your Experiment Web References PAM Sequences Glossary Publications CRISPR Overview...mutation within the targeted gene. However, the strength of the knockout phenotype for a given mutant cell... into high fidelity Cas9s (hfCas9) . There is currently no defined measure of fidelity, so sometimes they...pyrimidine-to-pyrimidine) Base transversions — edit to different classification of nucleotide base (purine-to-pyrimidine...genetic changes, including moving entire genes to different locations, by combining other gene editing mechanisms...
  10. Rett Syndrome

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    ...information, prenatal LMD microarray data, ISH image data, and anatomic reference atlases of prenatal and adult...approximately 1:10,000. It is characterized by apparently normal development through the first 6 months... skills occurs. Diagnosis of Rett syndrome is currently based on specific clinical diagnostic criteria... known to cause Rett syndrome, however, eight recurrent missense and nonsense mutations (R106W, R133C,...system for generating Rett syndrome-specific differentiated cells that can be used to understand the mechanisms...immortalized pre-neuronal cell line that can be differentiated into neurons Coriell Institute: Patient-specific...(Link opens in a new window) Rettsyndrome.org References Amir et al. 1999. Rett syndrome is caused by ...
  11. Immunology Research Plasmids and Resources

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    ...PAQR2 ADM adrenomedullin AM ADM2 adrenomedullin 2 AM2, FLJ21135, dJ579N16.4 ADRB1 adrenergic, beta-1-,... MGC138246 GDF1 growth differentiation factor 1 - GDF10 growth differentiation factor 10 BMP-3b, BMP3B...BMP3B GDF11 growth differentiation factor 11 BMP-11, BMP11 GDF15 growth differentiation factor 15 GDF-15,... PTGFB GDF2 growth differentiation factor 2 BMP-9, BMP9 GDF3 growth differentiation factor 3 - GDF5 growth...growth differentiation factor 5 BMP14, CDMP1, LAP4, OS5, SYNS2 GDF6 growth differentiation factor 6 BMP13...MGC158101, SGM1 GDF7 growth differentiation factor 7 BMP12 GDF9 growth differentiation factor 9 - GDNF glial...PPH1, T-ALK GDF1 growth differentiation factor 1 - GDF10 growth differentiation factor 10 BMP-3b, BMP3B...
  12. Fluorescent Protein Guide: Biosensors

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    ...for a different analyte, a sensor for gene-specific activity, or for popular biosensors in different expression...the articles linked below include variants with different binding affinities or negative-control constructs...3;32(40):13819-40. Baljit Khakh , Douglas Kim , Loren Looger , GENIE Project Calcium GCaMP3 genetically...indicators. Nat Methods. 2009 Dec;6(12):875-81. Loren Looger Calcium NEMO calcium sensors Engineering ...integrators. Science. 2015 Feb 13;347(6223):755-60. Loren Looger , Eric Schreiter Calcium Intersectional viral...cGMP (cyclic GMP) FlincG GFP-based cGMP sensor Differential patterning of cGMP in vascular smooth muscle...chloride sensitivity, photostability and reduced ph interference demonstrates augmented transmembrane chloride...
  13. Zhang Lab CRISPR Page

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    ...plasmid expressing SpCas9 alone without sgRNA. Full references are below. 1. SpCas9 (or SpCas9n, D10A nickase...Library CRISPR page for more information. Full references are below. Return to top Cas9 SAM transcriptional...The MS2-P65-HSF1 activation helper protein Full references are below. Pooled genome-wide human SAM library...Swiech et al., Nature Biotechnology 2014). Full references are below. Available plasmids are described below...The vector can be digested using BsaI ( This is different from BbsI, which is used in the SpCas9 plasmid...with Cas9 in a wide range of applications. Full references are below. The Cas9 knockin mouse can be purchased...This plasmid contains two expression cassettes, Renilla luciferase-2A-Cre recombinase and sgRNAs targeting...
  14. Optogenetics AAV Preps

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    ...pAAV-CaMKIIa-ChRmine-mScarlet-Kv2.1-WPRE CamKII ChRmine (high-photocurrent, red-shifted) mScarlet Constitutive 8 Karl Deisseroth...pAAV-hSyn-ChRmine-mScarlet-Kv2.1-WPRE Syn ChRmine (high-photocurrent, red-shifted) mScarlet Constitutive 8 Karl Deisseroth...pAAV-Ef1a-DIO-ChRmine-mScarlet-WPRE EF1a ChRmine (high-photocurrent, red-shifted) mScarlet Cre dependent 1, 5 Karl... 6m-p2A-ChRmine-Kv2.1-WPRE Syn ChRmine (high-photocurrent, red-shifted) GCaMP6m Constitutive 8 Karl Deisseroth... pAAV-nEF-ChRmine-mScarlet nEF ChRmine (high-photocurrent, red-shifted) mScarlet Constitutive 8 Karl Deisseroth...pAAV-nEF-Con/Fon-ChRmine-oScarlet nEF ChRmine (high-photocurrent, red-shifted) oScarlet Cre and Flp dependent...-Coff/Fon-ChRmine-oScarlet nEF ChRmine (high-photocurrent, red-shifted) oScarlet Flp dependent 8 Karl ...
  15. Genetic Code Expansion

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    ...native UGA recognition. Farren Isaacs Return to top Other Resources and References GCE4All Research Center... directed evolution to charge the tRNA with a different amino acid. To achieve these modifications, two...coli p-acetyl-l-phenylalanine (pAcF) Bacterial TAG Farren Isaacs 73545 pEvol-pAcFRS.1.t1 pAcFRS.1.t1 E. coli...coli p-acetyl-l-phenylalanine (pAcF) Bacterial TAG Farren Isaacs 73546 pEvol-pAzFRS.2.t1 pAzFRS.2.t1 E. coli...coli p-azido-l-phenylalanine (pAzF) Bacterial TAG Farren Isaacs 73547 pEvol-pAzFRS.1.t1 pAzFRS.1.t1 E. coli...coli p-azido-l-phenylalanine (pAzF) Bacterial TAG Farren Isaacs 82417 pUltra-sY sY-specific aaRS M. jannaschii...EAziRS) p-Azido-phenylalanine (Azi) Mammalian TAG Irene Coin 105830 pNEU-hMbPylRS-4xU6M15 PylRS M. barkeri...
  16. Mammalian RNAi Tools

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    ...Overview of shRNA-mediated RNA interference. Created with BioRender.com. shRNA Plasmids Browse our selection...Plasmids for mammalian RNA interference (RNAi) experiments at Addgene; browse by constitutive lentiviral...Tools for RNAi Browse Plasmids Resources RNA interference (RNAi) is an RNA-mediated gene silencing mechanism...pSico with pSicoligomaker (Andrea Ventura lab) References Carthew, R. W., & Sontheimer, E. J. (2009). Origins...J. (2009). siRNA vs. shRNA: Similarities and differences . Advanced Drug Delivery Reviews, 61 (9), 746... Tuning silence: conditional systems for RNA interference . Nature Methods, 3 (9), 682–688. PubMed (Link...
  17. Fluorescent Protein Guide: Subcellular Localization

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    ...Beta-catenin GFP Adherens junctions beta-Catenin EGFP Alpha Yap 67937 mouse E-cadherin GFP (423) Adherens junctions...interest with a fluorescent protein that has a different emission wavelength. For example, if the marker...mGreenLantern Gregory Petsko 221898 mito-mStayGold_pTwistCMVBetaGlobWPRENeo Mitochondria Mitochondrial targeting...mGreenLantern Gregory Petsko 221899 mStayGold-aTubulin_pTwistCMVBetaGlobWPRENeo Microtubules alpha-Tubulin mStayGold...stress. There are four types of cell junctions: adherens junctions, gap junctions, tight junctions, and...EGFP Alpha Yap 71366 Murine E-cadherin mCherry Adherens junctions E-cadherin mCherry Alpha Yap 55114 mCherry-Paxillin...Michael Davidson 55001 mCherry-Beta-Catenin-20 Adherens junctions beta-Catenin mCherry* Michael Davidson...
  18. Rinehart Lab Phosphoprotein Reagents

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    ...Phosphoprotein Reagents Phosphoprotein Synthesis Reagents References & Protocols Additional Resources Figure 1: SepRS...amino acids on either side occurring within the parent protein. Using the recoded strain of E. coli , ...performing phosphoproteomic experiments in need of reference spectra or phosphopeptide standards for quantitative...The Mode #2 phosphosite library contains four different phosphobinding domains: 14-3-3β, 14-3-3σ, the ...phosphorylation positional isomers are allocated to different subpools, allowing precise knowledge of phosphorylated... SepOTSα SepRS/EF-Sep ( aka. pKD-SepRS-EFSep) References & Protocols Iterative Synthetically Phosphorylated...Umehara, T., Guo, L., Osborne, E. M., Benner, J., Noren, C. J., Rinehart, J., & Söll, D. (2011). Expanding...
  19. CRISPR Plasmids - Repress Gene Expression

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    ...Overview of CRISPR interference. Browse, sort, or search the tables below for CRISPR interference plasmids. To...expression by interfering with transcription. This interference is most efficient when the gRNA targets your... To learn more about CRISPR interference and other CRISPR topics, read our CRISPR Guide . Mammalian ID...list of CRISPR resources by visiting our CRISPR reference page . CRISPR Guide Viral Preps CRISPR Protocols...
  20. Tetracycline Inducible Expression

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    ...express both components in the same relative ratio. Different generations of transactivators and promoters are...new window) for comparison of Tet-On systems in different applications. Use of Tetracycline or a Derivative...172115 PB-TO-hNGN2 Piggybac Tet-On plasmid for differentiating iPSCs into glutamatergic neurons via human ...NGN2 into AAVS1 safe harbor site, for iPSC differentiation into cortical neurons Michael Ward 100798 pAAV-syn-FLEX-splitTVA-EGFP-tTA...Inducible Promoters Blog: Repressible Promoters References Das, A. T., Tenenbaum, L., & Berkhout, B. (2016...window) Krueger, C., Pfleiderer, K., Hillen, W., & Berens, C. (2004). Tetracycline derivatives: alternative...
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