... CRISPR References & Information Genomic Deletions in Mammalian ...elements in mammalian cell lines. Protocol For referenced Tables and Figures, please access the manuscript...(100 μM; see step 1.4 ), 1.0 μl 10x T4 Ligation Buffer, 6.5 μl ddH 2 O, and 0.5 μl T4 Polynucleotide Kinase...μM; see step 3.1.4 ), 5.0 μl restriction enzyme buffer (10x), 4.0 μl (20 U) BbsI restriction enzyme (5,000...successfully adapted to numerous cell types using preferred culture conditions and transfection strategies... before attempting genome editing. Immediately transfer solution from cuvette into 1 ml of culture media...Minimize the time between electroporation and transferring the solution into media to enhance cell viability...

...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 ...pOpen-NcoIM M.NcoI DNA methyltransferase 165520 pOpen-T4 PNK T4 PNK polynucleotide kinase 165521 pOpen-lambda red...red operon 165530 pOpen-T4-BGT T4-BGT beta-glucosyltransferase 165531 pOpen-UDG UDG Uracil DNA glycosylase...

...listing of transfer status for viral preps.... Viral Vector Packaging Service Penn Vector Core Transfer Viral ...Viral Vector Packaging Service: Penn Vector Core Transfer The University of Pennsylvania Vector Core (Link...inventory , which includes all viral vectors transferred from the Penn Vector Core. Chemogenetics (including...mRuby2-GSG-P2A-GCaMP6s-WPRE-pA Biosensor Tobias Bonhoeffer, Mark Huebener, Tobias Rose AV-1-50943 50943-...mRuby2-GSG-P2A-GCaMP6f-WPRE-pA Biosensor Tobias Bonhoeffer, Mark Huebener, Tobias Rose AV-1-68717 68717-...pAAV-CAG-Flex-mRuby2-GSG-P2A-GCaMP6s-WPRE-pA Biosensor Tobias Bonhoeffer, Mark Huebener, Tobias Rose AV-1-68719 68719-...

...γ-retroviruses, commonly referred to as retroviruses, are the most popular virus for gene transfer in clinical trials... is typically preferred for in-vivo studies because of its low immunogenicity. Different types of viruses...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... host. Use this guide to learn more about the different types of viruses that are commonly used in research...

...metabolite levels. Different plasmids and, often times, different bacterial strains have different levels of precision...pTac GFP, Luciferase (LuxCDABE) Attila Karsi Broad host range vectors for expressing luciferase and fluorescent...produce a therapeutic compound, there are many different ways to alter bacterial genomes. From recombineering...cleavage site. Includes sites for LIC cloning, and a "stuffer" fragment that includes the SacB gene, allowing...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...Quorum sensing molecule) Cynthia Collins Contains luciferase inducible by the quorum sensing molecule, 3OC6HSL...

...to LacZ, plus luciferase-2A-Cre recombinase 60226 : sgRNA cloning backbone with luciferase-2A-Cre recombinase...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...

...PhyBdeltaCreC and PIF6CreN, in AAV donor/transfer vector Cbh AAV Eisenhoffer 131786 pLenti-CreLite CreLite system...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...fusion, or expression system of choice. We also offer ready-to-use viral preparations of select Cre expression...backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR Cre, luciferase, and sgRNA expression EFS AAV Zhang 60229 AAV...Retroviral Luikart 67503 pF CAG luc-EGFP-cre puro Luciferase - EGFP- Cre fusion CAG Mammalian Stringer 68448...terminus Cre fused to PIF6 CMV Mammalian, Zebrafish Eisenhoffer 131781 pCS2+PhyBΔCreC CreLite - Red light-inducible...

...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 ...start of my target sequence? The human U6 promoter prefers a 'G' at the transcription start site to have high...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...

...TARDBP CMV ALS Shawn Ferguson 133754 pLVX-Puro-TDP-43-A90V TARDBP CMV ALS Shawn Ferguson 133755 pLVX-Puro-...TARDBP CMV ALS Shawn Ferguson 133756 pLVX-Puro-TDP-43-A382T TARDBP CMV ALS Shawn Ferguson 133757 pLVX-Puro-TDP...TARDBP CMV ALS Shawn Ferguson 133758 pLVX-Puro-TDP-43-K263E TARDBP CMV ALS Shawn Ferguson 133759 pLVX-Puro-TDP...TARDBP CMV ALS Shawn Ferguson 133760 pLVX-Puro-TDP-43-P112H TARDBP CMV ALS Shawn Ferguson 133761 pLVX-Puro-TDP...TARDBP CMV ALS Shawn Ferguson 133762 px459-TDP-43 sgRNA1 TARDBP U6 ALS Shawn Ferguson 133763 px459-TDP 43...Suzanne Pfeffer 186013 pCMV Flag-LRRK2 K17A/K18A/R1441G LRRK2 Flag CMV Parkinson's Suzanne Pfeffer 186016...Suzanne Pfeffer 186017 pQE-80L 2xHis-LRRK2 Armadillo 1-552 LRRK2 His T7 Parkinson's Suzanne Pfeffer 186018...

...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...as shRNA (short hairpin RNA) or siRNA (short interfering RNA). Delivery can occur via transfection or ...using viral vectors. Lentiviral vectors are a preferred method for long-term gene silencing. Once the ...below. Figure 1: Overview of shRNA-mediated RNA interference. Created with BioRender.com. shRNA Plasmids ...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...

...for a different analyte, a sensor for gene-specific activity, or for popular biosensors in different expression...You may also like... Addgene Blog: Biosensors Luciferase Plasmids Subcellular Localization Optogenetics...fluorescent protein (cpFP) or Förster resonance energy transfer (FRET) efficiency of a pair of fluorescent proteins...the articles linked below include variants with different binding affinities or negative-control constructs...environmental factors and the sensor behavior may differ from the original characterization (e.g. in vitro...the plasmids associated with the article. We also offer ready-to-use AAV preparations of select plasmids...Cre-dependent) Cell-specific restoration of stimulus preference after monocular deprivation in the visual cortex...

...formulation buffer is phosphate-buffered saline containing 1 mM sodium azide. This buffer is not compatible...columns. Formulation Buffer After affinity chromatography, antibodies undergo a buffer exchange using centrifugal... cells and will interfere with conjugation. For applications such as these, the buffer can be removed ...webpage. Antibody preparations developed at IPI may differ from Addgene antibodies in “Production & Usage”...extracted from cells, separated by SDS-PAGE, transferred to a membrane, and labeled with the antibody ...

...lentiviral and adenoviral transfer vectors with Addgene. Lentiviral Transfer Vectors Within this set of...packaging these transfer vectors and generating infectious virus. Protocol: Lentivirus Transfer Vector Production...Plasmid FAQs Biosafety Resource Guide Lentiviral Transfer Vectors Adenoviral Vectors Kazuhiro Oka and the... of lentiviral transfer vectors you can find reporter plasmids to test whether or not you've efficiently...coexpressed with tdTomato and Pac (puromycin N-acethyl-transferase) pCDH-CB-copGFP-T2A-iCre 72256 Expresses copGFP...PGK promoter Adenoviral Vectors The adenoviral transfer vectors contained within this set are derived ...Flp recombinases. Genes can be cloned into these transfer vectors using shuttle vectors pICPIS-EF1 and pICPIS-CB...

...Desired Genetic Manipulation Different genetic manipulations require different CRISPR components. Selecting...be present in a single lentiviral transfer vector or separate transfer vectors May contain reporter gene...experimental goal, you are ready to start navigating the different reagents that are available for your particular...particular cell type or organism without a specific preferred mutation Cas9 (or Cas9 nickase) Single (or dual...can make a limited set of mutations. Repress or Interfere (CRISPRi) Reduce expression of a particular gene...targeting promoter elements of target gene Many different activators exist, including the multi-plasmid ...transduction CRISPR elements are inserted into an AAV transfer vector and used to generate AAV particles (for...

...Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing...vector, but their function (cut, nick, activate, interfere, etc.) is limited to that of the Cas protein present...have plasmids with Cas9 that can cut, activate, interfere, or nick. Selection , such as Puromycin or EGFP...pyogenes EGFP Ebert pLKO.1-puro U6 sgRNA BfuAI stuffer 50920 Mammalian/Lentiviral BfuAI none S. pyogenes...Joung pCRISPathBrick 65006 Bacteria BsaI yes, interfere S. pyogenes Koffas BPK764 65767 Bacteria BsaI ...pyogenes Joung MSP712 65768 Bacteria BsaI yes, interfere S. pyogenes Joung sgRNA with U6 promoter 48962...pyogenes Bar Chen pBUN6I11 50580 Plant BsaI yes, interfere S. pyogenes Bar Chen pBUN411 50581 Plant BsaI ...

...Kasparov Popular γ-Retroviral Transfer Plasmids Common γ-retroviral transfer plasmids. Want a retroviral...18760 MSCV IRES Luciferase MSCV Plasmid for transgene expression; also expresses luciferase Lowe 9044 pMIG...Addgene, including popular packaging, envelope, and transfer plasmids. Viral...scientists discuss retrovirus, they typically are referring to a subset of retroviruses called γ-retroviruses...of interest? Search our database for retroviral transfer plasmids that have inserts. ID Plasmid 5'LTR Description...Sun 60683 pLXIN-Luc MoMSV Stable expression of luciferase in mammalian cells Wong Return to Top Do you ...

...formulated in one of the following three buffers. All buffers are supplemented with 0.001% pluronic F68... or 0.014% Tween 20. Phosphate Buffered Saline (PBS) A universal buffer suitable for any application Balanced...as blood, vitreous, CSF, etc. Not suitable as a buffer for vectors delivered directly to the brain parenchyma...Shannon Boye’s lab at the University of Florida. References for each serotype can be found below under Citations...is available in each of the above serotypes and buffers. Please click on the plasmid name for ordering ...re Looking For? Our Packaged on Request service offers you even more options. Select from a vast catalog...

...cortical neuron differentiation. Held et al. Cell Rep. 2023 Aug 30. Enhance differentiation to mesencephalic...Collection CRISPR Tools AAV Viral Preps iPSC Differentiation Factors Antibodies Fluorescent Proteins Other...Amyotrophic lateral sclerosis (ALS), sometimes referred to as Lou Gehrig’s disease, is a progressive disease...are prone to misfolding and aggregation and can interfere with protein-protein interactions and nerve cell...of these are due to a mutation in one of three different inherited genes: Presenilin 1, Presenilin 2, and...al. Nat Methods. 2023 May 4. See More iPSC Differentiation Factors Find plasmids used to transform iPSCs...progression of Huntington’s disease. The foundation offers curated information on tools and reagents (Link...

...expressing firefly luciferase. pCMV-FLuc - Retroviral reporter vector expressing firefly luciferase. pHAGE-CMV-Luc2... vector expressing firefly luciferase and ZsGreen. NL4-3 mCherry Luciferase - Lentiviral dual reporter...EGFP. Lenti-luciferase-P2A-Neo - Lentiviral reporter vector expressing firefly luciferase with neomycin...reporter vector expressing mCherry and firefly luciferase. pLentiEGFPdestablized - EFS-EGFPd2PEST-2A-MCS-Hygro...

... mutations in methyl-CpG-binding protein 2 confer different severity in Rett syndrome. Neurology . 70,...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... editing of mutations in the MECP2 gene. Please refer to Addgene's CRISPR Guide for a general introduction...microarray data, ISH image data, and anatomic reference atlases of prenatal and adult human brain (Link...(Link opens in a new window) Rettsyndrome.org References Amir et al. 1999. Rett syndrome is caused by ...MeCP2 levels display tissue- and cell-specific differences and correlate with neuronal maturation. Hum Mol...