...), 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...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...been included. Ensure there are 2 x 10 6 cells per CRISPR pair. Resuspend 2 x 10 6 cells in 100 μl of electroporation... each primer pair designed based on testing the bulk sorted cells. Run samples on 2% agarose gel at 10...Deletions in Mammalian Cell Lines via CRISPR/Cas9. Bauer DE, Canver MC, Orkin SH. J. Vis. Exp. (95), e52118, ...frequency of indels formed on non-deleted alleles 2 , which are likely to cause frameshift mutations leading...of the Pim1 sgRNA from Table 1 are found in Table 2 . Obtain 24- or 25-mer oligos for each guide and its...

...proximal origin of SARS-CoV-2 . BSG - Wang, K et al, 2020, BioRxiv SARS-CoV-2 invades host cells via a novel...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...targeting the SARS-CoV-2 nucleocapsid protein (Brian Geiss). Plasmids SARS-CoV-2 Plasmids Many plasmids...packaging plasmids. SARS-CoV-2 Lentiviral preps now available for several SARS-CoV-2 constructs from the Krogan...SARS-CoV-2 plasmids are curated on a separate Ginkgo Bioworks Plasmid Collection page. SARS-CoV-2 Pooled...

...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,...HPS, HPS2, PE AZGP1 alpha-2-glycoprotein 1, zinc-binding ZA2G, ZAG B2M beta-2-microglobulin - CALR calreticulin...

...pseudotyping with SARS-CoV-2 spike protein.... 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... express ACE2, the cellular receptor for SARS-CoV-2, in a single round of viral entry and infection. This...particles have been pseudotyped with the SARS-CoV-2 Spike protein and you will find examples of each in...

...SARS-CoV-2 Plasmids The following types of plasmids are available: Bacterial expression: SARS-CoV-2 genes...speed up research of SARS-CoV-2, Ginkgo Bioworks is using their DNA design and synthesis capabilities, ...Browse plasmids encoding SARS-CoV-2 viral proteins. Depositor... that will enable researchers to express SARS-CoV-2 viral proteins in several expression systems. Addgene...expression and/or purification Yeast expression: SARS-CoV-2 genes in yeast expression vectors with various C-terminal...and/or purification Mammalian expression: SARS-CoV-2 genes in mammalian expression vectors with various...expression and/or purification Truncated SARS-CoV-2 Spike gene constructs for making VSV pseudovirus Common...

...Name Description PI 141391 pLVX-EF1alpha-SARS-CoV-2-N-2xStrep-IRES-Puro Expression of SARS-CoV-2 nucleocapsid...141386 pLVX-EF1alpha-SARS-CoV-2-M-2xStrep-IRES-Puro Expression of SARS-CoV-2 membrane (M) protein Krogan...141385 pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro Expression of SARS-CoV-2 envelope (E) protein Krogan...141383 pLVX-EF1alpha-SARS-CoV-2-orf3a-2xStrep-IRES-Puro Expression of SARS-CoV-2 orf3a protein Krogan 141347...Barcode Library Version 2 Ready-to-use lentiviral particles carrying version 2 of the CellTag barcoding...barcoding library. Version 2 of the Celltag library contains 4934 barcodes to combinatorially index cells for...pBOB-CAG-SARS-CoV2-Spike-HA Expression of SARS-CoV-2 spike (S) protein Pao 154985 pWPI-IRES-Puro-Ak-ACE2...

...guide RNA (gRNA) for biochemical purification (Fig. 2). For additional information and protocols, check ... web page . Figure 1: iChIP general scheme Figure 2: enChIP general scheme Fujii Lab Plasmids Individual... of interest 85586 gRNA Cloning Vector Bbs I ver. 2 An empty sgRNA expression vector. sgRNA sequence can... into BbsI site. gRNA scaffold has efficient ver. 2 structure. 92220 pLenti_dCas9-2xAM Lentiviral plasmid...Stedingk K, Fujita T, Persson CU, Eriksson P, Veerla S, De Preter K, Speleman F, Fujii H, Påhlman S, Mohlin ...Mohlin S. Biochem Biophys Res Commun. 2018 May 5;499(2):291-298. doi: 10.1016/j.bbrc.2018.03.150. PubMed . ...also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts Originally developed 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...Photosynthesis-dependent H 2 O 2 transfer from chloroplasts to nuclei provides a high-light signalling mechanism...Calcium Red fluorescent calcium sensors (fRCaMP1/2, GECO1/2) The kinetic mechanisms of fast-decay red-fluorescent...intracellular Zn(2+) homeostasis. Nat Methods. 2009 Oct;6(10):737-40. Maarten Merkx Zinc eZinCh-2 Zn2+ FRET ... eZinCh-2: A Versatile, Genetically Encoded FRET Sensor for Cytosolic and Intraorganelle Zn(2+) Imaging...

...FTZ-F1beta, LRH-1, LRH1, hB1F-2 nuclear receptor subfamily 5, group A, member 2 NR6A1 Plasmids NR6A1, GCNF...which interacts with hormones or other molecules, and 2) a DNA binding domain, which targets the receptor ...ESR-BETA,ESRB, ESTRB, Erb, NR3A2 estrogen receptor 2 (ER beta) ESRRA Plasmids ESRRA, ERR1, ERRa, ERRalpha...-1R nuclear receptor subfamily 1, group D, member 2 NR1H3 Plasmids NR1H3, LXR-a, LXRA, RLD-1 nuclear receptor...429G5.1, TLL, TLX, XTLL nuclear receptor subfamily 2, group E, member 1 NR2F1 plasmids NR2F1, COUP-TFI,...SVP44, TCFCOUP1, TFCOUP1 nuclear receptor subfamily 2, group F, member 1 NR3C1 Plasmids NR3C1, GCCR, GCR...NR3C2VIT nuclear receptor subfamily 3, group C, member 2 NR4A1 Plasmids NR4A1, GFRP1, HMR, N10, NAK-1, NGFIB...

...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...phosphorylation-dependent protein-peptide interactions (“mode #2” expression, Barber et al, Nat. Biotech. 2018). The...fluorescence-activated cell sorting (FACS). The mode #2 phosphosite library with four different phosphobinding... 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...Knockout Human Doench 3rd 2 40,964 Ingolia lab S cerevisiae CRISPRi v1 – barcodes 181005 161769 Inhibition...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...

...expression of gRNA under control of the Drosophila U6:2 promoter Bullock 49410 pCFD3-dU6:3gRNA expression ...CRISPR Fly Design CRISPR Fly Design You may also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR...please refer to the CRISPR Fly Design homepage. gRNA plasmids ID Name Description PI 49408 pCFD1-dU6:1gRNA ...CRISPR Fly Design is an open science genome engineering project in Drosophila. Genome... CRISPR Blog Posts Background CRISPR fly design is a genome engineering project in Drosophila, consisting...multiplex gRNA plasmid Bullock Cas9 plasmids ID Name Description PI 62209 pAct-Cas9 Expresses Cas9 under control...

...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...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...

...SpCas9n with 2a-Puro and 2a-EGFP are also available. 2. SpCas9 (or SpCas9n, D10A nickase) + CRISPR RNA array... system - lentiCRISPR - sgRNA and SpCas9 together 2 vector system - lentiCas9-Blast and lentiGuide-Puro...two MS2 RNA aptamers at the tetraloop and stemloop 2 The MS2-P65-HSF1 activation helper protein Full references...backbone with MS2 loops at tetraloop and stemloop 2 and EF1a-zeo resistance marker. Contains BsmBI sites...inverted terminal repeats (ITR) from AAV serotype 2. SaCas9 only: This plasmid ( PX600, #61592 ) contains...Epub 2013 Aug 29. Erratum in: Cell. 2013 Oct 10;155(2):479-80. PubMed . Genome engineering using the CRISPR-Cas9..., Scott DA, Mikkelsen TS, Heckl D, Ebert BL, Root DE, Doench JG, Zhang F. Science . 2014 Jan 3;343(6166...

...mouse somatic cells. Cell Stem Cell. 2008 Feb 7. 2(2):151-9. Jaenisch Lentivirus Mouse Polycistronic, ... cell reprogramming factors and wait for cells to de-differentiate. However it may be difficult to decide...Nature of Induced Pluripotency. Cell. 2015 Jul 16;162(2):412-24. Mikkelsen Lentivirus Human Expression of ...self-replicative RNA. Cell Stem Cell. 2013 Aug 1;13(2):246-54. Dowdy Adenovirus Mouse Non-integrating expression...reprogramming in mouse. Cell Stem Cell. 2008 Mar 6. 2(3):230-40. Hochedlinger MMLV-derived Retrovirus Mouse...Lentiviral Human Small molecules enable neurogenin 2 to efficiently convert human fibroblasts into cholinergic...

... mutations in the gene methyl-CpG binding protein 2 ( MECP2 ). MECP2 Rett syndrome is an X-linked disorder...in transcriptional repression, activity dependent de-repression, chromatin remodeling, gene activation...X-linked MECP2, encoding methyl-CpG-binding protein 2. Nat Genet . 23, 185–188. (Link opens in a new window... Cuddapah et al. 2014. Methyl-CpG-binding protein 2 (MECP2) mutation type is associated with disease severity... Specific mutations in methyl-CpG-binding protein 2 confer different severity in Rett syndrome. Neurology...males with mutations in Methyl-CpG binding protein 2. Am J Med Genet B Neuropsychiatr Genet . 180, 55–67...species (mouse or human) Mutation ID Plasmid Description Gene/Insert Mutations PI Return to top External...

...Can I order more than 2 µg of cloning grade DNA? At this time, to order more than 2 µg of a single plasmid...feedback on this service. DNA Service Details Amount 2 µg Volume 20 µl Concentration 100 ng/µl (± 5ng/µl)...Plasmids Available as Cloning Grade DNA ID Plasmid Description Industry PI FAQ What Quality Control analyses...cloning grade DNA into cells? Cloning grade DNA is designed for use in cloning applications. Direct transfection...

...Bacterial TAG Thomas Huber 73544 pEvol-pAcFRS.2.t1 pAcFRS.2.t1 E. coli p-acetyl-l-phenylalanine (pAcF) Bacterial...Bacterial TAG Farren Isaacs 73546 pEvol-pAzFRS.2.t1 pAzFRS.2.t1 E. coli p-azido-l-phenylalanine (pAzF) Bacterial...TAG Huiwang Ai 71403 pCMV-DnpK PylRS M. barkeri N6‐(2‐(2,4‐dinitrophenyl)acetyl)lysine (DnpK) Bacterial,...DiZSeK, or DiZHSeC Mammalian TAG P. Chen 173897 SepRS(2)/pSertRNA(B4)/EF-Sep aminoacyl-tRNA synthetase M. ...Mm-PylRS-AF/Pyl-tRNACUA PylRS M. mazei trans-cyclooct- 2-ene-lysine (TCOK) Mammalian TAG Howard Hang 163915...Huber 174718 pRSF-G1mCNPRS G1mCNPRS M. archaeon L-3-(2-cyano-4-pyridyl)alanine (mCNP) Bacterial TAG Thomas...Huber 174719 pRSF-G1pCNPRS G1pCNPRS M. archaeon L-3-(2-cyano-5-pyridyl)alanine (pCNP) Bacterial TAG Thomas...

...pAAV.CMV.PI.EGFP.WPRE.bGH James M. Wilson AV-2-27056 27056-AAV2 pAAV-Ef1a-DIO EYFP Karl Deisseroth AV-2-ALL854 51502-AAV2 AAV... Hongkui Zeng AV-2-PV0101 105530-AAV2 pAAV.CMV.PI.EGFP.WPRE.bGH James M. Wilson AV-2-PV0101 105530-AAV2...James M. Wilson AV-2-PV1090 105537-AAV2 pENN.AAV.CMVs.Pl.Cre.rBG James M. Wilson AV-2-PV2004 105545-AAV2...Wilson AV-2-26967M 100049-AAV2 pAAV.hSynap.ChETA(E123T/H134R)-eYFP.WPRE.hGH Karl Deisseroth AV-2-35505 35505...EYFP Karl Deisseroth AV-2-PV0102 105531-AAV2 pAAV.CMV.LacZ.bGH James M. Wilson AV-2-PV1917 105541-AAV2 pENN.AAV.CamKII0.4...AAV2 pAAV.CMV.PI.EGFP.WPRE.bGH James M. Wilson AV-2-PV1963 105542-AAV2 pENN.AAV.CB7.CI.eGFP.WPRE.rBG James...PV3447 59171-AAV1 pAAV-Syn-ChrimsonR-tdT Ed Boyden AV-2-26969P 26969-AAV2 pAAV-CaMKIIa-hChR2(H134R)-EYFP Karl...

... cause up to 2~3-fold repress on targeted fluorescent genes. These plasmids are described in: Repurposing...could cause up to 300-fold repress on targeted genes. 2. Two-plasmid CRISPRi system for mammalian gene knockdown...promoter and sgRNA targeting GFP (NT1) 46917 pU6-sgCXCR4-2 Human pSico-based U6 vector containing murine U6 promoter... UAS promoter (negative control) 46918 pU6-sgCD71-2 Human pSico-based U6 vector containing murine U6 promoter...also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts We have recently repurposed...protein and a customized s ingle g uide RNA (sgRNA) designed with a 20-basepair complementary region to any...ampicillin resistance, wherein the N20 can be custom designed to target arbitrary sequences in the genome. Co-expression...