... Temperature Genetically encoded ratiometric fluorescent thermometer A highly-sensitive genetically encoded...sensors for metabolites like fructose-1,6-bisphosphate , cyclic di-GMP , or S-adenosyl methionine . Reporters...Fluorescent Biosensors Fluorescent Proteins: Biosensors Metal Ions Small Molecules Neurotransmitters Cell Physiology...; these are noted with the viral service icon . Metal Ions Target Description Article PI Calcium jGCaMP8...neuronal populations and microcompartments. Nat Methods. 2019 Jul;16(7):649-657. Douglas Kim , GENIE Project... with large dynamics and high sensitivity. Nat Methods. 2023 Jun;20(6):918-924. Yubin Zhou Calcium Twitch...bioRxiv 2023.10.12.562058. Robert Campbell Calcium Ratiometric calcium biosensors from nested reporters of green...

...Lindquist 15589 MET FLAG46QDProCFPp303 HTT FLAG, CFP MET1 Huntington's Susan Lindquist 15590 MET FLAG72QDProCFPp303...FLAG, CFP MET1 Huntington's Susan Lindquist 15591 MET FLAG103QDProCFPp303 HTT FLAG, CFP MET1 Huntington's...FTD Ahmet Yildiz 159719 pOmnibac_ZZ_linker_ybbR_Tau MTBD MAPT ZZ, ybbR polH Parkinson's, FTD Ahmet Yildiz...Carlos Igual 89925 Profiline PFN1 T7 ALS Guillaume Romet-Lemonne 92200 pINDUCER20-delta-Syn SNCA Myc TRE2...

..., cell viability, and tumor metastasis. Currently, the most popular method for conducting genome-wide ... to conventional methods for genomic purification, CRISPR-based purification methods are more straightforward...1.1). Phage-assisted continuous evolution (PACE) methods resulted in xCas9 3.7 which has 7 mutations found...inducing double-strand breaks by modifying the methylation state of cytosines in a gene’s promoter or by...by inducing histone acetylation or demethylation. CRISPR epigenetic tools are specific for particular ... libraries containing Cas9 are the most popular method for CRISPR screening, they are not suitable for...hybridization (FISH), CRISPR imaging offers a unique method for detecting the chromatin dynamics in living ...

...difficult to decide which method to use. Addgene's blog post Delivery Methods for Generating iPSCs provides...combinations A more efficient method to generate integration-free human iPS cells. Nat Methods. 2011 May;8(5):409...an overview of several different reprogramming methods with the goal of helping readers choose a strategy...strategy suited to their research. Once you know which method you would like to use, browse Addgene's collection...cell into another via transdifferentiation. This method of cell lineage conversion can be faster, bypasses... stem cells (iPSCs). Sort the table by delivery method, species, or PI and then select the article link...link to find more information. Delivery Method Species Description Article PI Lentivirus Human Expression...

...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...

...Activation Human Zhang 3rd 10 96,458 Human CRISPR Metabolic Gene Knockout Library 110066 Knockout Human Sabatini...Human Vakoc 3rd 6 3,051 Human Lipid Droplet and Metabolism Library 191535 Knockout Human Olzmann 3rd ~10...Knockout Mouse Yusa 3rd 5 90,230 Mouse CRISPR Metabolic Gene Knockout Library 160129 Knockout Mouse Birsoy...169082 Knockout Mouse Turner N/A 10 13,500 Mouse Metastasis CRISPR-Cpf1(Cas12a) Double Knockout Library 123361...Inhibition Yeast Tavazoie N/A 6-12 51,681 Mouse Metabolism Library 163966 Knockout Mouse Kimmelman 3rd ~...Knockout Mouse X.S. Liu 3rd 5 5 10 49,252 Human Metabolic Gene CRISPRa sgRNA Library 187080 Activation Human... Human Birsoy 3rd Varies 32,460 Mouse Metabolic Gene CRISPRa sgRNA Library 187081 Activation Mouse Birsoy...

... expression in rEcoli XpS in the Methods section link below (Methods for iSPI, rEcoli XPS, pSerOTS-C1*...phosphoserine in >36,000 phosphosites by tandem mass spectrometry (Barber et al, Nat. Biotech. 2018). This phosphosite...human substrates of kinases of interest by mass spectrometry in a technique called serine-oriented library...resource described in (Gassaway et al., Nature Methods 2022) is adapted from the synthetic human phosphoproteome...phosphorylated positions for testing phosphoproteomics methods, proteomics search algorithms, proteomics phosphosite...phosphosite localization algorithms, or any method confounded by phosphosite ambiguity. Rinehart Lab Improved...Paulo JA, Rinehart J, Huttlin EL, Gygi SP. Nat Methods 2022 Nov;19(11):1371-1375. doi: 10.1038/s41592-...

... and 6.2 recommend an efficient method for DNA extraction, any method for genomic DNA isolation may be...and genetic elements. Here we present a simple methodology for CRISPR design, cloning, and delivery for ... Anneal in a thermocycler using the following parameters: 37 °C for 30 min; 95 °C for 5 min and then ramp...samples in a thermocycler using the following parameters: Cycles 1-20 (37 °C for 5 min, 20 °C for 5 min...system. NOTE: Alternatively use another transfection method such as cationic liposome-based transfection. Optimize...proliferated once macroscopically visible with ~2 mm diameter. Primer Validation and Screening for CRISPR/Cas9...samples in a thermocycler using the following parameters: 95 °C for 15 min, 35 cycles of (95 °C for 30...

...criteria have disease-causing mutations in the gene methyl-CpG binding protein 2 ( MECP2 ). MECP2 Rett syndrome...is not clear, however, the ability to bind to methylated DNA and recruit known co-repressors or other ...NID demonstrating the importance of binding to methylated DNA and the recruitment of the NCoR complex for...below: Base Editing RNA Editing CRISPR-based DNA methylation editing system - Shawn Liu and Rudolf Jaenisch...clinical trials (Link opens in a new window) HRV Metrics Toolbox - algorithms for data analysis to calculate...caused by mutations in X-linked MECP2, encoding methyl-CpG-binding protein 2. Nat Genet . 23, 185–188....new window) PMID: 16905679 Cuddapah et al. 2014. Methyl-CpG-binding protein 2 (MECP2) mutation type is ...

...Hang 163915 pRSF-ChPylTMSK ChPylTMSK N6-(((Trimethylsilyl)methoxy)carbonyl)-L-lysine (TMSK) Bacterial TAG...200226 pMega-MaFRSA PylRS M. alvus meta-trifluoromethylphenylalanine Bacterial TAG Alanna Schepartz 31186...David Tirrell 63177 pMarsL274G methionyl-tRNA synthetase (L274GMmMetRS) M. musculus azidonorleucine (Anl...ATG David Tirrell 89189 pMaRSC methionyl-tRNA synthetase (L274GMmMetRS) M. musculus azidonorleucine (Anl...TAA Wenshe Liu 85484 pDule-tfmF A65V S158A tri-fluoromethyl-phenylalanine synthetase M. jannaschii family...p-iodo-L-phenylalanine Bacterial TAG Dieter Söll 51401 pAM1 NLL-MetRS E. coli azi-donorleucine (Anl) Bacterial ATG David...

...(Spain). At this conference, mentors and mentees met and exchanged impressions of the IMP experience. ...with mentors who are working in USA and Europe, something that I would not able to do without a program ... particularly interested in breast cancer and metastasis as well as teaching and mentoring activities....

... to be successful, multiple requirements must be met. First, CRISPR would need to be delivered to both...respectively. Although each study used slightly different methods, they each observed the same positive phenotypes...

...Expression mAmetrine 406 526 26 6 0.8 hr Monomer (A206K) pBad-mAmetrine - Bacterial Expression mAmetrine-N1 -... these proteins, see: Cranfill et al. , Nature Methods 2016 Jul;13(7):557-62. FPbase : A database for ... - Mammalian Expression mAmetrine-C1 - Mammalian Expression LSSmOrange 437 572 23 5.7 2.3 hr Monomer pLSSmOrange-C1...Jump to Top References: Cranfill et al. : Nature Methods July 2016, Vol. 13(7):557-62 Olympus Confocal :...December 2007, pp. 4247-4260 Shaner et al. : Nature Methods, Vol.5 No. 6, June 2008, pp. 545-51 Shcherbakova.... 24 No. 5, pp.577-81 Shcherbo et al. : Nature Methods, September 2007, Vol. 4 No. 9, pp. 741-6 Shemiakina...2005, Vol. 10 No. 1, p. 14003 Chu et al. : Nature Methods, May 2014 Vol. 11 No. 5, pp.572-8 Yu et al. : Nature...

...lysate method. Purity Purity of AAV preparations is assayed by comparing the relative stoichiometric ratios...Overview of Addgene's viral production methods and quality control for lentivirus and AAV. Viral...Production Viruses are generated using standard methods that have been optimized for each specific virus...Details about our production protocols, titering methods, and quality control are described below. AAV Production...higher than those achieved using the standard qPCR method. Titering by the University of Pennsylvania Vector...gene expression and/or function. These data are sometimes reported or posted on the material page for the...

...including histone acetylation/demethylation, and cytosine methylation/demethylation....p300 histone demethylation by LSD1 cytosine methylation by DNMT3A or MQ1 cytosine demethylation by Tet1 These...

...TALE fusions with the histone demethylase LSD1 (Lysine specific demethylase) made using these plasmids along...with the FLASH method can decrease the amount of Histone H3 lysine 4 (H3K4) di-methylation at the target...Ekker Golden Gate TALEN 2.0 Golden Gate cloning method. Validated in multiple organisms. Dan Voytas Adam... Zhang Lab TALE Toolbox PCR/Golden Gate cloning method. Optimized for human expression. Feng Zhang LIC...(assembled using the REAL, REAL-Fast, or FLASH methods) for expression as TALE-LSD1 fusions. It was recently...

...FLASH assembly methods can be found on the TALengineering website . All three of these methods yield TAL effector...arrays assembled using the REAL, REAL-Fast or FLASH methods. TALengineering... (assembled using the REAL, REAL-Fast or FLASH methods) can be cloned for expression as TALE transcriptional...artificial transcription factors ( Maeder et al., Nat Methods 2013 ). All expression vectors listed below have... of the assembly are different among the three methods, the final products are the same). The publicly...

...Ligation) assembly method (Sander et al., Nat Biotechnol. 2011), the REAL-Fast assembly method (Reyon et al....effectors, including designed TALENs, and assembly methods for REAL, REAL-Fast and FLASH Genome...Automatable Solid-phase High-throughput) assembly method (Reyon and Tsai et al., Nat Biotechnol 2012). All...architectural framework and the DNAs produced by all three methods are identical in sequence. TAL effector repeat ... activators in human cells (Maeder et al., Nat Methods 2013) More detailed information about REAL, REAL-Fast...

...the Materials and Methods of your future publications. For your Materials and Methods section: "The plasmid...Plamids for Oligomerized Pool ENgineering (OPEN) method has been used to engineer ZFNs that function in...ZFNs via the Oligomerized Pool ENgineering (OPEN) method. This kit consists of 5 individually packaged plasmids...2009 Sep 17. PubMed . Description Use of the OPEN method requires OPEN pools (available by request from ...2000 ). The Oligomerized Pool ENgineering (OPEN) method has been used to engineer ZFNs that function in...

...are initially titered by spectrophotometry using a NanoDrop spectrophotometer. If needed, the sample is...Overview of Addgene's antibody production methods and quality control processes for purified recombinant... scientists. They are generated using standard methods individually optimized for each specific antibody...Details about our production protocols, titering methods, and quality control are described below. Information...PCR product is purified, sequenced via the Sanger method, and aligned with the parental plasmid sequence...