...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, ...

...Taipale 87065 pcDNA3.1-ccdB-Renilla Renilla Creation of C-terminal Renilla luciferase fusions using Gateway...Mikko Taipale 87068 pcDNA3.1-Renilla-ccdB Renilla Creation of N-terminal Renilla luciferase fusions using ...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...luciferase Brent Stringer 102320 FUW-RLuc-T2A-PuroR Renilla Ubc Lentiviral Expression of renilla luciferase...Diego Orzaez 68215 pEGB 35S:Renilla:Tnos (GB0109) Renilla 35S Expression of Renilla luciferase in plants ...12179 pIS1 Renilla Insertion of target sequence in the 3' UTR David Bartel 12177 pIS2 Renilla Insertion...

...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...

...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...

...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...

...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...

...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...

...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 ...

...yeast Gal1 Yeast Sandmeyer 26888 pmCherry-CRY2-CreN CRY2-CreN and mCherry coexpression; Light inducible; ...Constructs Cre Reporters & Tools Additional Resources References Background The Cre-lox system, derived from P1...respectively) and placed under the control of different promoters. Expression of both N and CCre in the...high-titer viral preparations of plasmids that are currently in our repository. ID Plasmid Description Promoter... Roxed-Cre GFAP Mammalian Pelczar 51267 pCAG-Co-InCreN N-terminal component of the Co-InCre system CAG... cre recombinase CAG AAV Bruchas 75268 pAAV-CRY2-CreN photo-activatable cre recombinase CAG AAV Bruchas...Cre-pA Cre; for atrial-specific expression ANF AAV Wehrens 121675 pAAV-EF1a-fDIO-Cre Cre EF-1 alpha AAV Engel...

...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...Cre-dependent) Cell-specific restoration of stimulus preference after monocular deprivation in the visual cortex...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...

...I Use? Use this guide to learn more about the different types of viruses that are commonly used in research... cancer therapy because of their ability to preferentially infect and kill cancer cells. Adenoviruses ...(through standard cloning methods). There are different types of viruses that are commonly used for research...research, each of which exhibit different properties, and thus, are suited for specific research goals...in-vivo studies because of its low immunogenicity. Different types of viruses also vary in the composition ... host. Use this guide to learn more about the different types of viruses that are commonly used in research...provides a brief overview of the properties of different types of viruses. More detailed information can...

...tools to perform different types of genome targeting. They are very good source of references. For genome ...double nickase system is that you can express two different chimeric gRNAs with the Cas9 nickase which will... like the wildtype Cas9 does. There are a few references for the double nickase system, including one ...system probably have better specificity. A good reference is this Cell paper: http://www.cell.com/abstract...most guides would work, but they sometimes have different efficiency). And it is best if the cut site is...for one well in 24-well plate. A good example/reference is this Cell paper: http://www.cell.com/abstract...vector, for a one well in a 24-well plate. A good reference is this paper in Cell . Click here for a 2013 ...

...Deletion Isolate gDNA from parental and bulk sorted cells by resuspending parental and bulk cell pellets in... CRISPR References & Information Genomic Deletions in Mammalian ...elements in mammalian cell lines. Protocol For referenced Tables and Figures, please access the manuscript... cells are suspension cells, instructions for adherent cells have also been included. Ensure there are.... NOTE: This protocol is for suspension cells. Adherent cells can either grow as individual cells in 96...concentration exceeds ~100,000 cells/ml or for adherent cells, the cells have reached ~80% confluence....Continue to step 7.3 for suspension cells. For adherent cells, aspirate media. Add 20 μl of 0.05% trypsin-EDTA...

...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...

...metabolite levels. Different plasmids and, often times, different bacterial strains have different levels of precision...produce a therapeutic compound, there are many different ways to alter bacterial genomes. From recombineering... pRSETa mEos2 20341 mEos2 (fluorescent protein) Loren Looger Photoactivable Fluorescent Protein Expression...protein expression. Vectors come with and without different combinations of His and Strep tags as well as ...control of gene expression so be sure to try a few different systems with your gene/protein of interest. Plasmid...to 3OC6HSL. EsaR derivatives found herein have different sensitivities to 3OC6HSL. The EsaR-responsive ...site compatible with standard cloning using many different restriction enzymes. pPro24-gfp contains GFP but...

...pAAV-CaMKIIa-ChRmine-mScarlet-Kv2.1-WPRE CamKII ChRmine (high-photocurrent, red-shifted) mScarlet Constitutive 8 Deisseroth...pAAV-hSyn-ChRmine-mScarlet-Kv2.1-WPRE Syn ChRmine (high-photocurrent, red-shifted) mScarlet Constitutive 8 Deisseroth...pAAV-Ef1a-DIO-ChRmine-mScarlet-WPRE EF1a ChRmine (high-photocurrent, red-shifted) mScarlet Cre dependent 1, 5 Deisseroth... 6m-p2A-ChRmine-Kv2.1-WPRE Syn ChRmine (high-photocurrent, red-shifted) GCaMP6m Constitutive 8 Deisseroth... pAAV-nEF-ChRmine-mScarlet nEF ChRmine (high-photocurrent, red-shifted) mScarlet Constitutive 8 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 Deisseroth...

...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...

... directed evolution to charge the tRNA with a different amino acid. They are typically expressed from ...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...-like click amino acids, tRNA M15 Mammalian TAG Irene Coin 113644 pRF0G-Tyr tyrosyl-tRNA synthetase M....

...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...FACS). The mode #2 phosphosite library with four different phosphobinding domains (14-3-3β, 14-3-3σ, the ...phosphorylation positional isomers are allocated to different subpools, allowing precise knowledge of phosphorylated...Library 111708 Mode #2 Library (NEDD4-2 WW2 domain) References & Protocols Iterative Synthetically Phosphorylated...Hohn MJ, Umehara T, Guo LT, Osborne EM, Benner J, Noren CJ, Rinehart J, Söll D. Science . 2011 Aug 26;333...

...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...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 54219 mEmerald-Paxillin...Michael Davidson 55001 mCherry-Beta-Catenin-20 Adherens Junctions Beta-Catenin mCherry Michael Davidson... 24920 pEGFP-LC3 (human) Autophagosome LC3 EGFP Toren Finkel 14435 mRFP-Clc Clathrin-coated vesicles Clathrin...