...10676 pMKO.1 GFP MoMLV U6-driven plasmid for shRNA expression; also expresses GFP. See pMKO.1 puro for ...reverse transcriptase and integrase. Once in a target cell, the virus uses the reverse transcriptase in tow...long-term transgene expression. Some disadvantages of γ-retroviruses are that they can only transduce dividing...Kasparov Popular γ-Retroviral Transfer Plasmids Common γ-retroviral transfer plasmids. Want a retroviral...Plasmid for transgene expression; also expresses luciferase Lowe 9044 pMIG MSCV Plasmid for transgene expression...Addgene, including popular packaging, envelope, and transfer plasmids. Viral...envelope). Additionally, γ-retroviruses integrate randomly into the host's genome, which can lead to oncogenesis...

...element Gordon Fishell 83900-AAV1 pAAV-mDlx-GFP-Fishell-1 GFP expression in forebrain GABA-ergic interneurons...element Gordon Fishell 83900-AAV2 pAAV-mDlx-GFP-Fishell-1 GFP expression in forebrain GABA-ergic interneurons...element Gordon Fishell 83900-AAV5 pAAV-mDlx-GFP-Fishell-1 GFP expression in forebrain GABA-ergic interneurons...element Gordon Fishell 83900-AAV8 pAAV-mDlx-GFP-Fishell-1 GFP expression in forebrain GABA-ergic interneurons...element Gordon Fishell 83900-AAV9 pAAV-mDlx-GFP-Fishell-1 GFP expression in forebrain GABA-ergic interneurons... Gordon Fishell 83900-AAVrg pAAV-mDlx-GFP-Fishell-1 GFP expression in forebrain GABA-ergic interneurons... Gordon Fishell 83900-PHPeB pAAV-mDlx-GFP-Fishell-1 GFP expression in forebrain GABA-ergic interneurons...

...Lentiviral BsmBI yes, cut S. pyogenes EGFP Ebert pLKO.1-puro U6 sgRNA BfuAI stuffer 50920 Mammalian/Lentiviral...Worm HindIII none S. pyogenes de Bono sgRNA with rpr-1 promoter 48961 Worm EcoRI none S. pyogenes de Bono...Zebrafish BseRI none S. pyogenes Zon pCRISPomyces-1 61736 Bacteria BbsI yes, cut S. pyogenes Apramycin...see plasmid page yes, cut S. pyogenes BFP Kuhn pLKO.1-puro U6 sgRNA BfuAI large stuffer 52628 Mammalian/...Puro Zoldos pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110) 71707 Mammalian hU6 yes, cut S. pyogenes Draetta...expression of Cas9, dCas9, or dCas9-VP64 along with 1-4 sgRNAs expressed from independent promoters. sgRNAs...for the expression of two gRNAs from Drosophila U6:1 and U6:3 promoters. Bullock pCFD5 Drosophila Utilizes...

...phage display. J Immunol Methods. 2000 Aug 28;242(1-2):159-81. Carlos Barbas Vector system for expression...immunostaining. Biochem Biophys Res Commun. 2015 Jan 2;456(1):527-33. Gavin Wright pET-30-based vector dedicated...mAb Collection . Once the plasmids are transiently transfected into mammalian tissue culture cells, the...making synthetic nanobodies (sybodies) against membrane protein targets. Synthetic single domain antibodies...antibodies for the conformational trapping of membrane proteins. Elife. 2018 May 24;7. and Generation of...

...populations defined by multiple parameters. Figure 1: Examples of intersectional cell population definitions... as a proof-of-concept targeting approach in 2014 1 (using EYFP and ChR2-EYFP as payloads). This approach...interneurons encode fear memory. Nat Neurosci. 23(1):61-74. PubMed (Link opens in a new window) Hafner...Itch-Scratching Cycle via Descending Regulation. Neuron 101(1):45-59. PubMed (Link opens in a new window) Lazaridis...including dual-transgenic recombinase-expressing mouse lines 2–9 and combinations of transgenic recombinase-expressing... Cre AND Flp, here expressed through a double transgenic animal strategy (PV-2a-Cre; SOM-IRES-Flp). Image...in a new window) of Flp-recombinase-expressing transgenic mouse and rat lines. Plasmids In addition to ...

... regulate a genome instead of modifying a genome. 1. Two-plasmid CRISPRi system for bacterial gene knockdown...repetitive sequence of human MUC4 exon 3 46915 pU6-sgGAL4-1 Human pSico-based U6 vector containing murine U6 promoter...efficiently block transcription (in bacteria, ~300-fold repression) by interfering with transcriptional elongation...for selection or fluorescent gating of transiently transfected cells. Co-expression of both plasmids in...elongation, RNA polymerase binding, or transcription factor binding. The binding specificity is determined jointly...the nuclease-deficient mutant could silence transcription on the gene expression level without genetically...CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes. Gilbert LA, Larson MH, Morsut...

...Bacterial TAG Farren Isaacs 73545 pEvol-pAcFRS.1.t1 pAcFRS.1.t1 E. coli p-acetyl-l-phenylalanine (pAcF) Bacterial...Bacterial TAG Farren Isaacs 73547 pEvol-pAzFRS.1.t1 pAzFRS.1.t1 E. coli p-azido-l-phenylalanine (pAzF) Bacterial...7FTrp) Bacterial TAG Thomas Huber 207639 pEVOL-NBK-1 PylRS M. mazei Pyrrolysine Bacterial Cole DeForest...and tyrU gene replaced by GentR) and release factor 1. Expresses archaeal MjTyrRS/tRNA pair instead. For...protein translation machinery. To expand the genetic code, 4 major changes to the standard translation machinery...Bacterial Ahmed Badran 209739 pAB228i24 G1 PylRS Methanogenic archaeon Bacterial Ahmed Badran 209740 pAB228q12...jannaschii Bacterial Ahmed Badran 209741 pAB228r3 Mlum1RS Bacterial Ahmed Badran 209742 pAB228p PylRS M. ...

...donor and acceptor. FRET is often used to study (1) protein-protein interactions where each protein is...standard consisting of ECFP and EYFP connected by 1 flexible glycine- and serine-containing peptide linker...fused to the N-terminus of LSSmOrange pLSSmOrange-C1 Orange Mammalian Express a gene of interest fused ...resonance energy transfer, or FRET , is a process by which energy is non-radiatively transferred from an excited... GFP variant) commonly used with mRuby2 pLSSmOrange-N1 Orange Mammalian Express a gene of interest fused...excited donor fluorophore to an acceptor. Since the transfer of energy does not occur by emission of a photon...FRET experiments. The efficiency of the energy transfer is measured using fluorescence microscopy by exciting...

...sequences containing a G nucleotide at position 20 (1 bp upstream of the PAM) may be more efficacious than...population. Some cells may be wild type due to either (1) a lack of gRNA and/or Cas9 expression or (2) a lack... in vitro and in vivo. Nature Biotechnology . 33 (1), 73–80. PMID: 25357182...For easy-to-transfect cell types (e.g. HEK293 cells), transfection with standard transfection reagents may...mammalian cell line that can be transfected at high efficiency Lentiviral transduction Cas9 and gRNA can be present...present in a single lentiviral transfer vector or separate transfer vectors May contain reporter gene ...screens using CRISPR AAV transduction CRISPR elements are inserted into an AAV transfer vector and used to ...

...for any application TMN200 (200 mM sodium chloride, 1 mM magnesium chloride, 20 mM Tris at pH 8.0) Suitable...adeno-associated viral vector. Hum Gene Ther Methods . Feb;25(1):72-82. PMID: 24191859 Ye, et al. 2015. Safety and...enhanced transduction of the retina following subretinal administration and enhanced transduction and spread...cite Boye, et al. 2016. Impact of Heparan Sulfate Binding on Transduction of Retina by Recombinant Adeno-... from the University of Florida for enhanced transduction of the eye and other tissues. As part of our...Y444F) serotype is demonstrated to have enhanced transduction of the retina following intravitreal injection... AAV2(Y444F) serotype also displays enhanced transduction of the trabecular meshwork following intracameral...

...Buenrostro JD, Regev A, Zhang F. Cell 2023 Jan 5; 186(1):209-229. PubMed (Link opens in a new window) . TF...Browse the Zhang lab’s comprehensive human transcription factor collection, available at Addgene. The...The collection can be used for screening transcription factors for cellular programming or cancer drug ...Library The MORF library is a comprehensive human transcription factor collection that contains 1,836 genes ...collection, can be purchased at Addgene. Each transcription factor isoform has a unique 24 bp barcode that...that simplifies identification of the transcription factor isoform in pooled screens. This collection ...collection and library can be used for screening transcription factors for applications such as cellular programming...

...in mammalian cells or zebrafish). pMG10 (a '“2-in-1” ZFN cassette for expression of both a “+” and “-”...different cell types. Zinc finger arrays can be transferred in-frame to these vectors. Plasmid...either OPEN or modular assembly can be easily transferred in-frame to any of these vectors using unique...

...Expression mEGFP 488 507 34 6 Monomer (A206K) pmEGFP-1 - Mammalian Expression mEGFP-N1 - Mammalian Expression...TurboRFP-pBAD - Bacterial Expression tdTomato 554 581 95 4.7 1 hr Tandem-dimer pCSCMV:tdTomato - Mammalian Expression...-pBAD - Bacterial Expression mRFP1 584 607 13 4.5 1 hr Monomer pcDNA3-mRFP - Mammalian Expression pMXs-mRFP1... of Biomedical Optics, Feburary 2005, Vol. 10 No. 1, p. 14003 Chu et al. : Nature Methods, May 2014 Vol... Monomer pET mOrange LIC cloning vector (u-mOrange) - Bacterial Expression pCS2+8CmOrange - Zebrafish/...pCS2+8NmOrange - Zebrafish/Xenopus/C.elegans/Sea urchin mOrange-N1 - Mammalian Expression mOrange-C1 -...Expression mOrange-pBAD - Bacterial Expression mOrange2 549 565 35 6.5 4.5 hr Monomer mOrange2-N1 - Mammalian...

...research and innovation programme and EFPIA. Figure 1: Representation of RESOLUTE mission. The combination...CRISPR knockout library targeting human transmembrane transporters. Feedback Please send your feedback to...carrier proteins (SLCs) are the largest family of transporters in the human genome with approximately 450 identified... Medicines Initiative 2 Joint Undertaking under grant agreement No 777372. This Joint Undertaking receives...allow a fast and efficient sub-cloning into a wide range of Gateway-compatible pDEST vectors. ID Plasmid ...overexpression library of human SLC proteins. Human Transporter Knockout Libraries 213695 221409 221410 Superti-Furga...

...Biology, 2(9), 537–541. DOI: 10.1021/sb400021j Figure 1: Multiplexing CRISPR knockdown to modulate expression...controlling the number of mismatches we can obtain a range of expression levels. This collection comprises ...Together, these plasmids allow to explore the whole range of expression levels for these two reporters simultaneously... S. (2018). Tuning dCas9’s ability to block transcription enables robust, noiseless knockdown of bacterial...

...gene activation CRISPR/Cas9 Plasmid Variants Figure 1: Schematic of the CRISPR Collection. Plasmids 159654... and 159655 are single guide expressing double stranded break constructs. Plasmids 171098 and 175570 allow...

...plasmids can be ordered via the links below: Table 1. dCas9 Activators sgRNA Dual Expression ID Plasmid...CRISPR Blog Posts CRISPR-on is an RNA-guided transcriptional activator system with two components, the dCas9...sgRNA). While the dCas9 protein provides the transactivation activity, the sgRNA component provides the ...dCas9 activators and sgRNAs can be separately transfected (pmax dCas9 activators [48223~48227] + sgRNA ...Gateway donor vectors [48218~48221] and can be transferred to Gateway destination vectors (e.g., pmax-DEST...endogenous genes by CRISPR-on, an RNA-guided transcriptional activator system. Cheng AW, Wang H, Yang H,...H, Shi L, Katz Y, Theunissen TW, Rangarajan S, Shivalila CS, Dadon DB, Jaenisch R. Cell Res. 2013 Oct;...

...proteins in budding yeast. Sci Rep. 2019 Feb 19;9(1):2234. doi: 10.1038/s41598-019-38913-z. PubMed 30783202...Looger LL, von Wirén N, Frommer WB. A cytosolic trans-activation domain essential for ammonium uptake....

...repression. The new transactivator rtTA ( r everse t etracycline-controlled t rans a ctivator) was created...CMV rtTA3 Blast (w756-1) Lentiviral reverse tetracycline-controlled transactivator 3 (rtTA3) expression...TetR On Geley Transactivators (tTA or rtTA) Find a construct that expresses the transactivator for your tetraycline...Overview of tetracycline regulated inducible transcriptional activation or repression Plasmid...tetracycline and not to the TRE, permitting transcription. Tetracycline-dependent promoters are developed... gene expression, a tetracycline-controlled transactivator (tTA) was developed. tTA was created by fusing... of VP16 (virion protein 16), an essential transcriptional activation domain from HSV (herpes simplex ...

...) a T ranscriptional R epressor D omain (TRD) the TRD contains a Nuclear receptor Co-Repressor 1/Silencing...affecting females at an incidence of approximately 1:10,000. It is characterized by apparently normal development...reactivation, RNA editing, gene replacement therapy, RNA trans-splicing, and protein replacement to reverse the...cases Rett syndrome can be inherited. In females, random X inactivation results in approximately half of...disease severity. The MECP2 protein is a global transcriptional regulator of thousands of genes and studies...studies have suggested roles in transcriptional repression, activity dependent de-repression, chromatin remodeling...DNA and recruit known co-repressors or other transcriptional modifiers suggests the major function of MECP2...