...Ensure there are 2 x 10 6 cells per CRISPR pair. Resuspend 2 x 10 6 cells in 100 μl of electroporation solution...Dilute oligos 1:10 in ddH 2 O ( e.g., 1.0 μl annealed oligos + 9.0 μl ddH 2 O to yield a concentration...), 50-100 ng gDNA, and H 2 O up to 20 μl. Use the primers designed in step 2 above. Conduct PCR for “non-deletion...supplemented with 2% penicillin/ streptomycin and 1% L-glutamine, which is used for MEL cells. However, transient...expression construct. Electroporate cells with 250 volts for 5 msec in a 2 mm cuvette using an electroporation...designed based on testing the bulk sorted cells. Run samples on 2% agarose gel at 10 V/cm using 1x Tris-acetate-EDTA...frequency of indels formed on non-deleted alleles 2 , which are likely to cause frameshift mutations leading...

...Transcription factor SOX-2 Transcription Factor 124607 ACTN2-mEGFP AICSDP-63 mEGFP Alpha-actinin-2 Sarcomeric z-disks...101781 CETN2-mTagRFP-T AICSDP-22 mTagRFP-T Centrin-2 Centrioles 101782 LAMP1-mEGFP AICSDP-19 mEGFP LAMP... AICSDP-83 mEGFP EZH2 Polycomb repressive complex 2 164500 POLR2A-mEGFP AICSDP-117 mEGFP RPB1 RNA polymerase...mEGFP AICSDP-77 mEGFP Telomeric repeat-binding factor 2 (TRF2) Telomeres 168799 CTCF-mEGFP AICSDP-144 mEGFP...for Cell Science has deposited plasmids to generate fluorescently tagged proteins of key cellular structures... Protein Guide Subcellular Localization Plasmids Protocols The Allen Institute for Cell Science has produced...fluorescently tagged proteins of key cellular structures in human cell lines of their choice. For a full ...

... reprogramming of mouse somatic cells. Cell Stem Cell. 2008 Feb 7. 2(2):151-9. Jaenisch Lentivirus Mouse...during fibroblast to iPS cell reprogramming in mouse. Cell Stem Cell. 2008 Mar 6. 2(3):230-40. Hochedlinger... a synthetic self-replicative RNA. Cell Stem Cell. 2013 Aug 1;13(2):246-54. Dowdy Adenovirus Mouse Non-integrating...Dynamic Nature of Induced Pluripotency. Cell. 2015 Jul 16;162(2):412-24. Mikkelsen Lentivirus Human Expression...Lentiviral Human Small molecules enable neurogenin 2 to efficiently convert human fibroblasts into cholinergic...Conversion of Fibroblasts. Neuron. 2014 Oct 22;84(2):311-23. Yoo Fibroblasts Neurons Lentiviral Human ...

...Durocher et al. Nucleic Acids Research 2002 Jan 15;30(2):E9 A list of commonly-used bait and prey plasmids...Consortium plasmids that encode the extracellular regions of cell surface receptor and secreted proteins...expression plasmids that encode the extracellular regions of cell surface receptor and secreted proteins...screens or cellular assays to identify novel interactions or pathways that are important in cellular recognition...AVEXIS (for Avidity-based extracellular interaction screen) in which the extracellular regions of receptor ...and Wright GJ Molecular & cellular proteomics 2010 Dec; 9(12):2654-65. A cell surface interaction network... Plasmid Collections EXPRESs Plasmids EXtracellular Plasmid RESource (EXPRESs) Consortium You may...

...proximal origin of SARS-CoV-2 . TMPRSS2 - Hoffmann M, et al, 2020, Cell SARS-CoV-2 Cell Entry Depends on ACE2...Converting Enzyme 2) is the host cell receptor mediating the entry of SARS-CoV and SARS-CoV-2 viruses. ( 1 ...that primes the SARS-CoV-2 S protein and is involved in virus entry into cells. ( 2 ) FURIN - an enzyme that...origin of SARS-CoV-2 . BSG - Wang, K et al, 2020, BioRxiv SARS-CoV-2 invades host cells via a novel route...page: SARS-CoV-2 Antibodies : Recombinant monoclonal antibodies that target SARS-CoV-2 nucleocapsid protein...protein. SARS-CoV-2 Plasmids : Plasmids that are available or coming soon containing SARS-CoV-2 sequences. SARS-CoV... targeting the SARS-CoV-2 nucleocapsid protein (Brian Geiss). Anti-SARS-CoV-2 Nucleocapsid Protein [mBG86...

...immunoglobulin heavy diversity 2-15 D2, IGHD215 IGHD2-2 immunoglobulin heavy diversity 2-2 IGHD22 IGHD2-21 immunoglobulin...heat shock 70kDa protein 2 HSP70-2, HSP70-3 HSPA4 heat shock 70kDa protein 4 APG-2, HS24/P52, MGC131852, ...variable 2-33 (non-functional) IGLV233, V1-9 IGLV2-8 immunoglobulin lambda variable 2-8 IGLV28, V1-2 IGLV3...beta 4 DEFB-2, DEFB102, DEFB2, HBD-2, SAP1 EDN1 endothelin 1 ET1, HDLCQ7 EDN2 endothelin 2 ET2, PPET2 ...receptor subfamily 2, group C, member 1 TR2 NR2C2 nuclear receptor subfamily 2, group C, member 2 TAK1, TR2R1...suppression of tumorigenicity 2 - STC1 stanniocalcin 1 STC STC2 stanniocalcin 2 STC-2, STCRP TAC1 tachykinin,... (natural killer cell stimulatory factor 2, cytotoxic lymphocyte maturation factor 2, p40) CLMF, CLMF2...

...transient mammalian expression 101126 pACP-tag (m)- 2 Vector ACP/MCP-tag Empty backbone for stable and transient... sub-cellular localization (membrane, nucleus, etc.) are also available Non-toxic to living cells To learn...repair protein, and is suitable for both cell-permeant and non-cell-permeant proteins. CLIP-tag CLIP-tag ...Control for cell surface localization 101131 pACP-GPI Control Plasmid ACP/MCP-tag Control for cell surface...visualization of proteins inside cells. Depositor... Depositor Collections New England Biolabs Cell-Imaging Plasmid Collection...Collection New England Biolabs Cell-Imaging Plasmid Collection Technology Plasmids Supplemental Information ...

...to infect permissive cells that express ACE2, the cellular receptor for SARS-CoV-2, in a single round of...pseudotyping with SARS-CoV-2 spike protein. Collections... Collections COVID-19 - Viral Pseudotyping SARS-CoV-2 Pseudotyped Virus...Virus Mechanisms of SARS-CoV-2 infection remain poorly understood but are critical for developing therapeutic...spread. As a Biosafety Level 3 (BSL-3) agent, SARS-CoV-2 requires specialized facilities for the study of the...replication-defective virus particles with the SARS-CoV-2 spike (S) glycoprotein on the envelope surface. Pseudotyped...particles have been pseudotyped with the SARS-CoV-2 Spike protein and you will find examples of each in...

...of basal H 2 O 2 levels with peroxiredoxin-based probes Real-time monitoring of basal H 2 O 2 levels with...Belousov Hydrogen Peroxide (H 2 O 2 ) Cytosolic or mitochondrial sensor for H 2 O 2 oxidation (roGFP2-Orp1) ...peroxiredoxin-2-based probe. Nat Commun. 2018 Aug 7;9(1):3145. Hadley Sikes Hydrogen Peroxide (H 2 O 2 ) Monitoring...Hydrogen Peroxide (H 2 O 2 ) HyPer biosensor to measure changes in intracellular hydrogen peroxide concentration...Hydrogen Peroxide (H 2 O 2 ) Expresses hydrogen peroxide biosensor HyPer2 in plant cells Photosynthesis-dependent...encoded Ca(2+) indicator with enhanced two-photon absorption. Neurophotonics. 2024 Apr;11(2):024207. doi...Calcium Red fluorescent calcium sensors (fRCaMP1/2, GECO1/2) The kinetic mechanisms of fast-decay red-fluorescent...

...status of MEK1/2 was tracked at 5 minute intervals via Western blot. Phosphorylation of MEK1/2 peaks at just...PubMed PMID: 28232577. Figures 2-4 were modified from this reference. 2. Regot, Sergi, Jacob J. Hughey...Treatment of the cells with a PP1/PP2A inhibitor leads to maintenance of phosphorylated MEK1/2 levels, suggesting...precisely control the timing of kinase activity. 2. Studying negative feedback loops of signaling pathways... studied the negative feedback on endogenous MEK1/2 phosphorylation by creating a short pulse of psRaf1...activity. They did this by exposing psRaf1 to a short 2 minute pulse of 500 nm light (turn on psRaf1) followed...suggesting PP1/PP2A is responsible for this MEK1/2 negative feedback. 3. Studying signaling pathways in...

...CCNB1 CCNB2 CCNB3 Cyclin B1, 2, or 3 Cyclin D CCND1 CCND2 CCND3 Cyclin D1, 2, or 3 Cyclin E CCNE1 CCNE2 ...kinase 1 CHK2 Checkpoint kinase 2 Cop-1 Ring finger and WD repeat domain 2 (RFWD2); E3 ubiquitin protein...SESN3 Sestrins 1, 2, or 3 Siah Siah E3 ubiquitin protein ligase 1 TSC2 Tuberous sclerosis 2 TSP1 Thrombospondin... Spring Harbor Perspectives in Biology. 2010 Feb;2(2):a001107. PMC PMID: 20182618 . Do you have suggestions...threonine kinase ATR ATR serine/threonine kinase B99 G-2 and S-phase expressed 1; also known as GTSE1 BAI-1...inhibitor 1A p48 Damage-specific DNA binding protein 2, 48kDa p53 Tumor protein p53 p53AIP1 Tumor protein... domain protein 1 PIGs Etoposide induced 2.4 PIRH-2 Ring finger and CHY zinc finger domain containing ...

...difficult to ignore [1, 2]. 2. Fusion to an obligate dimer or oligomer Several cell proteins normally self-associate.... J Cell Biol, 2003. 163(2): p. 257-69. PubMed PMID: 14581454. PubMed Central PMCID: PMC2173526. 2. Zacharias...Kd is unlikely to form dimers in a cell. The Kd of EGFP is 0.11 mM [2]. Following the simple logic outlined... nature are prone to dimerization (i.e. EGFP) [1, 2] or even form obligate tetramers (i.e. DsRed) [3]....organized smooth endoplasmic reticulum (OSER) whorls. 2)A dimerizing protein of interest fused to a dimerizing... designed to detect resulting in false positives [2, 4]. Solutions to the polymer problem There are some...Citrine) can be monomerized with an A206K mutation [2]. Addgene has many of these constructs and they are...

...same site, is 2’-fluoro (2’-F), which substitutes a fluorine for the hydroxyl group at the 2’ position. ...most common of these is 2’-O-methylation, where a methyl group is added to the 2’ hydroxyl group of the...them together, for example phosphorothioates with 2’-O-methyl modifications, proved more stability than...generated by introduction of a single photocleavable 2-nitrobenzyl linker in the 20-bp targeting region of...photocleavable gRNAs. Molecular Cell 81, 7, 1553-1565 (2021). https://doi.org/10.1016/j.molcel.2021.02.007 Moroz-Omori...evolved ways to prevent degradation of important cellular RNA molecules with post translational modifications...slightly more efficient and less toxic to mammalian cells (Allen, et al., Frontiers in Genome Editing).  Some...

...196682 kiCAP-AAV-M.Mus.2 M.Mus.2 Rep/Cap plasmid for the production of M.Mus.2, an AAV capsid with CNS...196688 kiCAP-AAV-M.Fas.2 M.Fas.2 Rep/Cap plasmid for the production of M.Fas.2, an AAV capsid with CNS...Expresses bicistronic AAV-2 rep gene and polycistronic AAV-2 cap gene in Sf9 insect cells Kotin 65215 pSR646 ...should be handled at BSL-2. AAV can transduce both dividing and non-dividing cells with a low immune response...Expresses bicistronic AAV-2 rep gene and polycistronic AAV-1 cap gene in Sf9 insect cells Kotin 65214 pSR657 ...Expresses bicistronic AAV-2 rep gene and polycistronic AAV-6 cap gene in Sf9 insect cells Kotin 65216 pSR660 ...Expresses bicistronic AAV-2 rep gene and polycistronic AAV-8 cap gene in Sf9 insect cells Kotin 92307 pAnc80L65AAP...

... on cell cycle) Centrin-2 EGFP Erich Nigg 41151 pEGFP Cep170 C-term Centrioles (dependent on cell cycle...sequence YFP Thorben Dammeyer 54520 mCherry-Peroxisomes-2 Peroxisome Peroximal Targeting Signal 1 (PTS1) mCherry...give the cell its shape, organize cellular organelles, and are necessary for movement and cell division...between animal cells. They are important for holding cell structures together and reducing cell stress. There... General Structures The cell contains a variety of organelles and subcellular structures each with their...example, houses the cell’s genetic material. The mitochondria produces the energy cells required for chemical...kappa-myc-dL5-2xG4S-TMst Cell surface (mammalian) PDGFR-derived dL5 FAP; mCerulean3 Marcel Bruchez 45944 pTDpelB-C_sfYFPTwinStrep...

...erythroid 2 like 2 NFKB1 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 PAK PAK1...E2F transcription factor ECT2 Epithelial cell transforming 2 EGFR Epidermal growth factor receptor EIF4EBP... Scribbled planar cell polarity protein SHC SHC1 SHC2 SHC3 SHC4 SHC (Src homology 2 domain containing)...ARHGEF2 Rho/Rac guanine nucleotide exchange factor 2 CCND CCND1 CCND2 CCND3 Cyclin D CDK CDK4 CDK6 Cyclin-dependent...enhancer of kinase suppressor of Ras CYTH2 Cytohesin 2 DUSP DUSP1 DUSP2 DUSP3 DUSP4 DUSP5 DUSP6 Dual specificity...binding protein ERBB2 Erb-b2 receptor tyrosine kinase 2 ERK MAPK1 MAPK3 Also known as MAPK; Mitogen-activated...homolog GRB2 Growth factor receptor bound protein 2 ICMT Isoprenylcysteine carboxyl methyltransferase ...

...fusion none 2, 5 Richie 121539 pAAV SYN1 HA-hM3D(Gq) hM3D(Gq) - Activation HA fusion none 2, 5 Richie 50467...none 2, 8 Roth 50476 pAAV-CaMKIIa-hM3D(Gq)-mCherry hM3D(Gq) - Activation mCherry fusion none 1, 2, 5, ...(Gq) - Activation mCherry fusion Cre-dependent 1, 2, 5, 8, 9, rg*, PHPeB Roth 44362 pAAV-hSyn-DIO-hM4D...(Gi) - Inhibition mCherry fusion Cre-dependent 1, 2, 5, 8, 9, rg*, PHPeB Roth 65417 pAAV-hSyn-dF-HA-KORD-IRES-mCitrine...mCherry hM3D(Gq) - Activation mCherry fusion none 1, 2, 5, 8, 9, rg* Roth 50475 pAAV-hSyn-hM4D(Gi)-mCherry...mCherry hM4D(Gi) - Inhibition mCherry fusion none 1, 2, 5, 8, 9, rg* Roth 52536 rAAV-CAG::FLEX-rev:: hM4D-...mCherry hM4D(Gi) - Inhibition mCherry fusion none 1, 2, 5, 8, 9 Roth 75033 pAAV CD68-hM4D(Gi)-mCherry hM4D...

...111707 Mode #2 Library (NEDD4 WW2 domain) Pooled Library 111708 Mode #2 Library (NEDD4-2 WW2 domain) References...Pooled Library 111705 Mode #2 Library (14-3-3β) Pooled Library 111706 Mode #2 Library (14-3-3σ) Pooled Library...discovered by fluorescence-activated cell sorting (FACS). The mode #2 phosphosite library with four different...phosphorylation-dependent protein-peptide interactions (“mode #2” expression, Barber et al, Nat. Biotech. 2018). The... WW2 domain of NEDD4, and the WW2 domain of NEDD4-2; 111705-8 ) is available from Addgene. Screening kinases...iSPI_pSer_Subpool#1 Pooled Library 188527 iSPI_pSer_Subpool#2 Pooled Library 188528 iSPI_pSer_Subpool#3 Pooled Library...pos ctrl Plasmid 111883 pNAS1b split mCherry NEDD4-2 WW2 Plasmid 111882 pNAS1b split mCherry NEDD4 WW2 ...

...3rd 1, 2, or 4 49,766 arrays Broad GPP kinome Brunello 75314, 75315 (1 plasmid) 75312, 75313 (2 plasmid...GPP genome-wide Brunello 73179 (1 plasmid) 73178 (2 plasmid) Knockout Human Doench and Root 3rd 4 76,441...Broad GPP genome-wide Brie 73632 (1 plasmid) 73633 (2 plasmid) Knockout Mouse Doench and Root 3rd 4 78,637...172651 (Set D) Knockout Human Doench and Root 2nd 2 40,710 Inzolia Human CRISPR/Cas12a Multiplex Knockout...3,052 Broad GPP kinome Brie 75317 (1 plasmid) 75316 (2 plasmid) Knockout Mouse Doench and Root 3rd 4 2,852...MinLibCas9 Library 164896 Knockout Human Garnett 3rd 2 37,722 Green monkey (Chlorocebus sabaeus) sgRNA library...Library (Gattinara) 136986 Knockout Human Doench 3rd 2 40,964 Ingolia lab S cerevisiae CRISPRi v1 – barcodes...

...expressing mCherry-tagged channelrhodopsin-2 driven by PV+ basket cell-targeting enhancer Gordon Fishell Viviana...expressing mCherry-tagged channelrhodopsin-2 driven by chandelier cell-targeting enhancer Gordon Fishell Viviana...construct expressing mCherry-tagged channelrhodopsin-2 driven by SST interneuron-targeting enhancer Gordon...construct expressing mCherry-tagged channelrhodopsin-2 driven by Lamp5 interneuron-targeting enhancer Gordon...construct expressing mCherry-tagged channelrhodopsin-2 driven by VIP interneuron-targeting enhancer Gordon...construct expressing mCherry-tagged channelrhodopsin-2 driven by cholinergic neuron-targeting enhancer Gordon...