...oligos 1:10 in ddH 2 O ( e.g., 1.0 μl annealed oligos + 9.0 μl ddH 2 O to yield a concentration of 1 μM)....35 cycles of (95 °C for 30 sec, 60 °C for 1 min, 72 °C for 1 min), and 72 °C for 10 min. Optimize PCR ...the page. CRISPR Design Design sgRNAs manually or using freely available online tools 1 . Use these tools...at the genomic recognition site. NOTE: Figure 1 describes possible deletion strategies for genes and non-coding...Use the example guides for the intended deletion of Pim1 in mouse ( Mus musculus ; Table 1 , Figure 2A )...Deletions in Mammalian Cell Lines via CRISPR/Cas9. Bauer DE, Canver MC, Orkin SH. J. Vis. Exp. (95), e52118, ...complement sequences of the Pim1 sgRNA from Table 1 are found in Table 2 . Obtain 24- or 25-mer oligos...

...gRNA-hIRF-1 #12 Expresses a guide RNA (gRNA) to target human IRF-1 promoter 61080 gRNA-hIRF-1 #12/pSIR ..._dCas9-2xAM_hIRF-1 Lentiviral plasmid expressing dCas9-2xAM and gRNA for human IRF-1 promoter. 98041 Sa-dCas9...interest. 98134 hIRF-1 gRNA #409 (Sa) Expresses a guide RNA (gRNA) to target human IRF-1 promoter for the ...system. 105283 hIRF-1 gRNA #351 (Sa) Expresses a guide RNA (gRNA) to target human IRF-1 promoter for the ...LexA are subjected to affinity purification (Fig. 1). In enChIP, specific genomic regions are tagged with...plasmids or browse the Fujii Lab web page . Figure 1: iChIP general scheme Figure 2: enChIP general scheme... Expresses a guide RNA (gRNA) to target human IRF-1 promoter 62190 3xFLAG-dCas9/pTEF1p-CYC1t Expresses...

...Cascade ( C RISPR- as sociated c omplex for a ntiviral de fense) complex, comprised of a combination of Cas5...endogenous repair mechanisms. Cas3 belongs to the Class 1 family of CRISPR systems, the most abundant type found...bacteria and archaea. While abundant in nature, Class 1 systems are largely underutilized compared to their...still separate the individual Cas components. Figure 1: Overview of Cascade-Cas3 mechanism. Created with ...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Cas3 is a unique member...

...IGHDY1 IGHD1-1 immunoglobulin heavy diversity 1-1 IGHD11 IGHD1-14 immunoglobulin heavy diversity 1-14 (non-...IGHV6-1 immunoglobulin heavy variable 6-1 IGHV61, VH IGHV7-4-1 immunoglobulin heavy variable 7-4-1 IGHV7... TAPBP-R, TAPBPR THBS1 thrombospondin 1 THBS, THBS-1, TSP, TSP-1, TSP1 TRPC4AP transient receptor potential...variable 2-8 IGLV28, V1-2 IGLV3-1 immunoglobulin lambda variable 3-1 IGLV31, V2-1 IGLV3-10 immunoglobulin lambda...NCC-1, NCC1, SCYA13, SCYL1 CCL14 chemokine (C-C motif) ligand 14 CC-1, CC-3, CKb1, FLJ16015, HCC-1, HCC... 3 G0S19-1, LD78ALPHA, MIP-1-alpha, MIP1A, SCYA3 CCL3L1 chemokine (C-C motif) ligand 3-like 1 464.2, D17S1718...GPD, GpFy, WBCQ1 DEFA1 defensin, alpha 1 DEF1, DEFA2, HNP-1, HP-1, MGC138393, MRS DEFA3 defensin, alpha...

...Function PI AV-1-49531P 100040-AAV1 pAAV.hSyn1.Twitch2B.WPRE.SV40 Biosensor Oliver Griesbeck AV-1-50942 50942...Tobias Rose AV-1-PV2723 98929-AAV1 pAAV.hSyn.iGluSnFr.WPRE.SV40 Biosensor Loren Looger AV-1-PV2724 98931... Looger AV-1-PV2725 98932-AAV1 pAAV.CAG.Flex.iGluSnFr.WPRE.SV40 Biosensor Loren Looger AV-1-PV2816 100835... Kim AV-1-PV2817 100839-AAV1 pAAV.CAG.Flex.GCaMP6m.WPRE.SV40 Biosensor GENIE, Douglas Kim AV-1-PV2818 ... Kim AV-1-PV2819 100833-AAV1 pAAV.Syn.Flex.GCaMP6f.WPRE.SV40 Biosensor GENIE, Douglas Kim AV-1-PV2820 ... Kim AV-1-PV2821 100845-AAV1 pAAV.Syn.Flex.GCaMP6s.WPRE.SV40 Biosensor GENIE, Douglas Kim AV-1-PV2822 ...Douglas Kim AV-1-PV2823 100841-AAV1 pAAV.Syn.GCaMP6m.WPRE.SV40 Biosensor GENIE, Douglas Kim AV-1-PV2824 100843...

...Constitutive 1, 9 Svoboda 83898 pAAV-mDlx-ChR2-mCherry-Fishell-3 Dlx ChR2 mCherry Constitutive 1, 9, rg* Fishell...Constitutive 1, 2, 5, 9 Deisseroth 26973 pAAV-hSyn-hChR2(H134R)-EYFP Syn ChR2/H134R EYFP Constitutive 1, 2, 5...Constitutive 1, 5, rg* Boyden 59171 pAAV-Syn-ChrimsonR-tdT Syn ChrimsonR tdTomato Constitutive 1, 5, 9 Boyden...dependent 1, 5, 9, rg* Deisseroth 26971 pAAV-CaMKIIa-eNpHR 3.0-EYFP CaMKII eNpHR 3.0 EYFP Constitutive 1, 9 ...Constitutive 1, 5, rg* Yizhar 125713 AAV-hSyn1-SIO-eOPN3-mScarlet-WPRE Syn eOPN3 mScarlet Cre dependent 1, 5 Yizhar...Constitutive 1, 5 Yizhar 198511 pAAV_hSyn-DIO-PdCO-mScarlet-WPRE Syn PdCO mScarlet Cre dependent 1, 5 Yizhar...Constitutive 1, 5 Yizhar 198516 pAAV_EF1a-DIO-PdCO-mScarlet-ER-miniWPRE EF1a PdCO mScarlet Cre dependent 1, 5 Yizhar...

..., hRev, THRA1, THRAL, ear-1 nuclear receptor subfamily 1, group D, member 1 NR1D2 Plasmids NR1D2, RVR,...subfamily 1, group D, member 2 NR1H3 Plasmids NR1H3, LXR-a, LXRA, RLD-1 nuclear receptor subfamily 1, group...AHX, DAX-1, DAX1, DSS, GTD, HHG, NROB1, SRXY2 nuclear receptor subfamily 0, group B, member 1 NR1D1 Plasmids...member 1 NR3C1 Plasmids NR3C1, GCCR, GCR, GR, GRL nuclear receptor subfamily 3, group C, member 1 (glucocorticoid...HMR, N10, NAK-1, NGFIB, NP10, NUR77, TR3 nuclear receptor subfamily 4, group A, member 1 NR4A2 Plasmids... FTZF1, POF7, SF-1, SF1, SPGF8, SRXY3 nuclear receptor subfamily 5, group A, member 1 NR5A2 Plasmids NR5A2...of the important regions of nuclear receptors are: 1) a ligand binding domain, which interacts with hormones...

...Fawzi 127194 RP1B FUS 1-163 QQ4xSS #1 FUS His T7 ALS Nicolas Fawzi 127195 RP1B FUS 1-163 QQ4xSS #2 FUS His... type 1 scFv [L24/1] ITPR1 CMV Spinocerebellar ataxia James Trimmer 206790 IP3 receptor, type 1 scFv [... His, V5 EF-1 alpha Parkinson's Mark Cookson 25082 pDEST51-LRRK2-R1441G LRRK2 His, V5 EF-1 alpha Parkinson's... His, V5 EF-1 alpha Parkinson's Mark Cookson 29399 pDEST51-LRRK2-I1371V LRRK2 His, V5 EF-1 alpha Parkinson's...pcDNA3-FLAG-Synaptojanin 1-145 SYNJ1 Flag CMV Parkinsonism, early-onset Pietro De Camilli 22292 pcDNA3-FLAG-Synaptojanin...-FLAG-Synaptojanin 1-170 SYNJ1 Flag CMV Parkinsonism, early-onset Pietro De Camilli 22293 pEGFPC1-FLAG-Synaptojanin...FLAG-Synaptojanin 1-145 SYNJ1 GFP, Flag CMV Parkinsonism, early-onset Pietro De Camilli 22294 pEGFPC1-FLAG-Synaptojanin...

...accumulation in bacteria. Biotechnol Biofuels. 2008 Jun 3. 1(1):11. Wolf Frommer Maltose Green fluorescent MBP-based...chloroplasts to nuclei provides a high-light signalling mechanism. Nat Commun. 2017 Jun 29;8(1):49. Philip Mullineaux...of cellular physiology. Nat Commun. 2020 Aug 4;11(1):3881. Adam Cohen Calcium GCaMP6f expression in forebrain...Dynamics in High-Ca Organelles. Cell Chem Biol. 2016 Jun 1. pii: S2451-9456(16)30163-5. Teresa Alonso , Javier... intracellular calcium. Nat Commun. 2021 Dec 9;12(1):7159. Dorus Gadella Calcium Teal genetically encoded...Neurophotonics. 2024 Apr;11(2):024207. doi: 10.1117/1.NPh.11.2.024207. Robert Campbell Calcium Ratiometric...orange-emitting fluorescent proteins. Nat Commun. 2017 Sep 5;8(1):431. Wolf Frommer Calcium Red-, yellow-, or cyan-...

...AdEasier®-1 cells (strain) - Bacterial strain that contains AdEasy®-1 plasmid...expression pMKO.1 puro - Retroviral shRNA expression pLenti CMV Puro DEST - ...Lentiviral Gateway destination vector for gene expression Hygromycin Mammalian pLKO.1 hygro - Lentiviral... expression Puromycin Mammalian pLKO.1 - TRC cloning vector - Lentiviral shRNA... - Retroviral gene expression pMKO.1 GFP - Retroviral shRNA expression pSico... expression, multiple selectable markers pMKO.1 puro - Mammalian retroviral shRNA expression...under your chosen promoter pAdEasy®-1 - Recombine plasmids from the shuttle...

... Human Doench 3rd 1, 2, or 4 49,766 arrays Broad GPP kinome Brunello 75314, 75315 (1 plasmid) 75312, 75313...Version 1 1000000069 Knockout Human Moffat 3rd 12 176,500 Toronto KnockOut - Version 3 90294 (1 plasmid...Library 182133 Knockout Human Mali 3rd 1 74 Perturb-seq Guide Barcodes (GBC) 85968 Barcode Human Weissman ...Libraries 153101-153106 Knockout Yeast Borodina N/A 1 Varies Bovine CRISPR Knockout Libraries 213927, 213928...pgRNAs 1,718 Broad 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...and Root 3rd 4 3,052 Broad GPP kinome Brie 75317 (1 plasmid) 75316 (2 plasmid) Knockout Mouse Doench and...

...Brown 11908 pEGFP-N1 alpha-actinin 1 Actin Filaments alpha-actinin 1 EGFP Carol Otey 27676 Cortactin-pmCherryC1...Huttenlocher 11908 pEGFP-N1 alpha-actinin 1 Focal Adhesions alpha-actinin 1 EGFP Carol Otey 31923 pPaxillin-PSmOrange...Autophagosome Sequestosome-1 EGFP Noboru Mizushima 38272 pMXs-IP GFP-WIPI-1 Autophagosome WIPI1 EGFP Noboru...apparatus eNOS(1-33) CFP Alexandra Newton 36205 pmTurquoise2-Golgi Golgi apparatus B4GALT1(1-61) mTurquoise2... Shirihai 158003 pCMV-mGold-CAF1-C-10 Nucleus CAF-1 mGold Francois St-Pierre 27705 CAV1-mCherry Caveolae...COXIV-COX8-dL5-2XG4S-mCer3 Mitochondria COX IV-derived (1-22 aa) import sequence and COX VIII signal peptide...Verkhusha 36208 pmTurquoise2-Mito Mitochondria COX8A(1-29) mTurquoise2 Dorus Gadella 26673 pEGFP-C1-Fibrillarin...

...Typical Titer Small 10 × 100 µL aliquots 1 mL 4 × 10 12 GC/mL* 1 × 10 13 GC/mL* Medium 25 × 100 µL aliquots...serotype (AAV1, AAV2, AAV5, AAV8, or AAVrg) and volume (1 mL, 2.5 mL, or 5 mL). Currently, we can only package...to 10 weeks. This breaks down as follows: Request 1–2 days Look for the banner on an eligible plasmid ...place your request. We will send you an email within 1-2 days with a response. Most requests will be approved... we can begin processing your order. MTA Approval 1+ business days, varies by requesting organization ...scientist through an MTA. This process typically takes 1–4 days, depending on your institution. AAV Production...all-in-one pricing for viral vector preps, which includes MTA facilitation, DNA amplification, and high-...

...positive feedback loop for amplified tTA expression. 1 Gradinaru 99121 pAAV-ihSyn1-DIO-tTA Cre-dependent ...positive feedback loop for amplified tTA expression. 1 Gradinaru 117383 TRE-DIO-eYFP Cre-dependent and Tetracycline-inducible...Tetracycline-inducible Cre-dependent, Tet-inducible expression of EYFP 1 Gradinaru Tools for Affinity Purification These AAV...synaptophysin-EGFP for labeling of axon terminals. 1 Zeng 71760 pAAV hSyn FLEx mGFP-2A-Synaptophysin-mRuby...synaptophysin-mRuby for labeling of axon terminals. 1 Luo 60658 pAAV-EF1α-F-FLEX-mNaChBac-T2A-tdTomato EF1a-driven...Cre-dependent Cre-dependent, bicistronic expression of designer pro-taCasp3 and TEVp for studying cell ablation...

...Cre-GFP fusion EF-1 alpha Mammalian Sauer 11923 pBS598 EF1alpha-EGFPcre Cre-EGFP fusion EF-1 alpha Mammalian...mCherry coexpression EF-1 alpha AAV Deisseroth 55635 pAAV-EF1a-sCre SCre EF-1 alpha AAV Deisseroth 55636...55636 pAAV-EF1a-Cre Cre EF-1 alpha AAV Deisseroth 55638 pAAV-EF1a-vCre VCre EF-1 alpha AAV Deisseroth 59701...) EF-1 alpha AAV Cepko 69571 pAAV-EF1a-C-CreintG Cre recombinase dependent on GFP (CRE-DOG) EF-1 alpha... Mammalian Sauer 11918 pBS513 EF1alpha-cre Cre EF-1 alpha Mammalian Sauer 11919 pBS448 RSV-GFPcre Cre-...11955 pBS505 EF1alpha-EGFPcre* Cre-EGFP fusion EF-1 alpha Mammalian Sauer 11956 pBS594 promoterless EGFPcre...GFAP-Cre Cre GFAP Mammalian Sofroniew 25997 LV-Cre pLKO.1 Cre and shRNA coexpression CMV Lentiviral Fuchs 26646...

...Institute ID Tag Protein Structure 87420 PXN-EGFP AICSDP-1 EGFP Paxillin Matrix Adhesions 87421 TUBA1B-mEGFP ...TJP1-mEGFP AICSDP-23 mEGFP Tight junction protein ZO-1 Tight junctions 91565 AAVS1-mEGFP AICSDP-35 mEGFP ...Centrioles 101782 LAMP1-mEGFP AICSDP-19 mEGFP LAMP-1 Lysosome 101783 MAP1LC3B-mEGFP AICSDP-25 mEGFP Autophagy-related...101786 ST6GAL1-mEGFP AICSDP-26 mEGFP Sialyltransferase 1 Golgi 109122 NPM1-mEGFP AICSDP-50 mEGFP Nucleophosmin... HIST1H2BJ-mEGFP AICSDP-52 mEGFP Histone H2B type 1-J Histones 109119 CTNNB1-mEGFP AICSDP-47 mEGFP Beta-catenin...Mitochondria 87424 DSP-mEGFP AICSDP-9 mEGFP Desmoplakin Desmosomes 87425 ACTB-mEGFP AICSDP-15 mEGFP Beta-actin...human cell lines of their choice. For a full description of how to use these plasmids, check out Plasmids...

...phosphosite library as GST-fusion peptides (“mode #1” expression). The mode #1 phosphosite library ( 111704...standards for quantitative assessments. The mode #1 phosphosite library can be generated with either phosphoserine...available from Addgene. Screening kinases. The mode #1 phosphosite library can also be synthesized using ...pSerOTS-C1* (V70) Pooled Library 188526 iSPI_pSer_Subpool#1 Pooled Library 188527 iSPI_pSer_Subpool#2 Pooled Library...split mCherry 14-3-3β Pooled Library 111704 Mode #1 Library Pooled Library 111705 Mode #2 Library (14-...partial UAG codon reassignment and release factor 1 deletion. Heinemann IU, Rovner AJ, Aerni HR, Rogulina...Serine Phosphopeptide Library described in: Encoding human serine phosphopeptides in bacteria for proteome-...

...plasmid Vogelstein 16399 AdEasier-1 BJ5183 derivative that contains AdEasy-1™ plasmid Vogelstein Adenoviral... that have inserts. ID Plasmid Type Description PI 16400 pAdEasy-1 Adenoviral For recombining shuttle ...for the AdenoBuilder genome assembly system. Block 1 with deletion in E1 and insertion of GFP expression...strains for generating adenovirus. ID Strain Description PI 16398 BJ5183 Strain for recombination between... be assembled in a one-tube reaction. ID Kit Description PI 1000000176 AdenoBuilder toolkit Plasmids contain...

... cell reprogramming factors and wait for cells to de-differentiate. However it may be difficult to decide... with CRISPR activators. Nat Commun. 2018 Jul 6;9(1):2643. Otonkoski Lentivirus Human Expression of human...cell reprogramming. Stem Cell Res Ther. 2017 Jun 5;8(1):132. Zovein Replicating EBNA1 episome Human Non-integrating...vector. Proc Natl Acad Sci U S A. 2009 Jan 6. 106(1):157-62. Jaenisch Lentivirus Mouse Doxycycline-inducible...transcription factors. Stem Cell Reports. 2015 Jan 13;4(1):25-36. Broccoli Fibroblasts Sensory Neurons Lentiviral...peripheral sensory neurons. Nat Neurosci. 2015 Jan;18(1):25-35. Baldwin Fibroblasts iTSCs Lentiviral Mouse... from Human ALS Patients. Cell Rep. 2016 Jan 5;14(1):115-28. Zhang Adult Dermal Fibroblasts Neurons Lentiviral...

...foreign DNA and cleaves it to destroy the invader ( Figure 1 ). Figure 1: An overview of the endogenous...separated by short palindromic repeat sequences. (1) The CRISPR array is transcribed to make the pre-CRISPR...engineering, with Type V following in 2015. Class 1 (Multi-subunit effector complex) Class 2 (Single multi-domain...enChIP) using CRISPR. Biochem Biophys Res Commun . 439(1):132-6. PMID: 23942116 Gilbert LA, Horlbeck MA, Adamson...X, Scott DA, Mikkelson T, Heckl D, Ebert BL, Root DE, Doench JG, Zhang F. 2014. Genome-Scale CRISPR-Cas9...CRISPR-Cpf1 using a single crRNA array. Nat Biotechnol . 35(1):31-34. PMID: 27918548...also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts The CRISPR revolution shows...