...Addgene’s pLKO.1 protocol. Pro-tip: this plasmid grows more slowly than standard plasmids. Find pLKO.1 - TRC...TRC cloning vector. Tet-pLKO-puro - Tet-pLKO-puro contains all the necessary elements for packaging, reverse... experiments.  Find the scrambled shRNA vector. pLKO.1 - TRC cloning vector - This vector from the David...target gene knockdown. Note: this plasmid was called pLKO-Tet-On in the original publication but its name ...manual provided by the Weiderschain lab. Find Tet-pLKO-puro. lentiGuide-Puro - This plasmid expresses the...

...backbones for shRNA expression, such as pLKO.1 puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids...Plasmids -  If you're familiar with the pLKO plasmids you'll note that those are lentiviral constructs. In...

...Recently I learned that Addgene’s pLKO.1 cloning protocol is viewed around 3,000 times each month. I ...protocol on the Addgene website in December 2006, the pLKO.1-TRC cloning vector (deposited by David Root of ...How would the thousands of other scientists using pLKO.1 variants learn about those updates? This is a problem...

...information and a detailed protocol on how to use the pLKO.1 vector can be found here. Cre-Lox technology Puro.Cre...here. Cloning and expressing new shRNA sequences pLKO (Plasmid #10878) This plasmid has been used by ...came about here. Cloning and expressing RNAi pMKO.1 puro (Plasmid #8452) RNA interference (RNAi) is an...

...expression pLKO.1 - TRC cloning vector - Lentiviral shRNA expression Tet-pLKO-puro - ... gene expression pLKO.1 hygro - Lentiviral shRNA expression Tet-pLKO-puro - ...activity shRNA/RNAi For gene silencing experiments pLKO.1 - TRC cloning vector - Lentiviral...AdEasier®-1 cells (strain) - Bacterial strain that contains AdEasy®-1 plasmid... - Retroviral gene expression Tet-pLKO-neo - Tet-inducible lentiviral shRNA... Tet-inducible lentiviral shRNA expression pMKO.1 puro - Retroviral shRNA expression ... - Retroviral gene expression pMKO.1 GFP - Retroviral shRNA expression pSico...

...for pMKO.1 puro; Contains shRNA against GFP William Hahn 10676 pMKO.1 GFP Derivative of pMKO.1 puro with...shRNA expression ID Plasmid Description PI 10878 pLKO.1 ‐ TRC cloning vector shRNA expression; Recommended...AgeI/EcoRI restriction digest) David Root 10879 pLKO.1 ‐ TRC control Negative control vector containing...containing non-hairpin insert David Root 1864 pLKO.1 ‐ scrambled shRNA Negative control vector containing scrambled...detailed protocol for using the popular cloning vector pLKO.1. Trono Lab Lentivectors Toolbox Tyler Jacks Lab...Description PI 8452 pMKO.1 puro Retroviral vector for shRNA expression Bob Weinberg 10675 pMKO.1 puro GFP shRNA ...role for Nramp1 in modifying susceptibility to type 1 diabetes. Kissler S, Stern P, Takahashi K, Hunter ...

...lenti". ID Plasmid Generation Description PI 8453 pLKO.1 puro 3rd U6-driven shRNA empty plasmid with puro...plasmid 24150 for hygro resistance. Weinberg 10878 pLKO.1 – TRC cloning plasmid 3rd U6-driven shRNA empty...; includes a stuffer for easy cloning Root 14748 pLKO.3G 3rd U6-driven shRNA empty plasmid with EGFP marker...for Thy1.1 selection. Benoist and Mathis 21915 Tet-pLKO-puro 3rd inducible expression of shRNA; puromycin...plasmid 21374 for dtomato version. Welm 21916 Tet-pLKO-neo 3rd inducible expression of shRNA; neomycin ...working with lentivirus: these viruses are based on HIV-1, which may require additional lab biosafety procedures... EGFP coexpression. Trono 17619 EF.CMV.RFP 2nd EF-1 alpha promoter for transgene and CMV drives expression...

...vectors (e.g. pBV-Luc) and gene silencing (e.g. pLKO.1 - TRC cloning vector). Check out our complete guide...

...Table of Contents A. pLKO.1-TRC Cloning Vector A.1 The RNAi Consortium A.2 Map of pLKO.1 A.3 Related plasmids...Order oligos compatible with pLKO.1 C. Cloning shRNA oligos into pLKO.1 C.1 Recommended materials C.2 Annealing...information Back to Top A. pLKO.1-TRC Cloning Vector A.1 The RNAi Consortium The pLKO.1 cloning vector is the...marker encoded in pLKO.1 allows for convenient stable selection. Figure 1 : Map of pLKO.1 containing an shRNA...Appendix I.1. Sequence of pLKO.1 TRC-Cloning Vector Click here to see the sequence of pLKO.1 TRC-cloning... Protocols pLKO.1 Protocol pLKO.1 - TRC Cloning Vector Addgene Plasmid 10878...arising from the use of this vector. A.2 Map of pLKO.1 pLKO.1 is a replication-incompetent lentiviral vector...

...and T4 polymerase pLKO.1 - TRC Cloning Vector Cloning protocols for using the pLKO.1 vector, a backbone...(PPE) for BSL-1 and BSL-2 Labs Learn how to best protect yourself when working in BSL-1 and BSL-2 labs...Levels One and Two (BSL-1 and BSL-2) Safety measures for laboratories operating at BSL-1 and BSL-2 Watch the...

...Lentiviral BsmBI yes, cut S. pyogenes EGFP Ebert pLKO.1-puro U6 sgRNA BfuAI stuffer 50920 Mammalian/Lentiviral... see plasmid page yes, cut S. pyogenes BFP Kuhn pLKO.1-puro U6 sgRNA BfuAI large stuffer 52628 Mammalian...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...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...

...Most shRNA‐expressing lentiviral plasmids, such as pLKO.1, use a U6 or H1 promoter in order to drive RNA ...detailed protocol for using the popular cloning vector pLKO.1. Trono Lab Resources: Lentivectors Toolbox and ...lentiviral transfer plasmids are based on the HIV-1 virus. For safety reasons, transfer plasmids are all...selection. Desfarges S, Ciuffi A. Viruses. 2010. Jan;2(1):111-30. PubMed . Altering the tropism of lentiviral...Curr Gene Ther. 2005. 5(4): 387-398. PubMed . HIV-1 Genome Nuclear Import Is Mediated by a Central DNA...

...Co-expressed tTA, rtTA, or TetR On or Off PI 21916 Tet-pLKO-neo 3rd generation lentiviral plasmid for inducible...Element On or off PI 26429 pLenti CMV rtTA3 Blast (w756-1) Lentiviral reverse tetracycline-controlled transactivator...

...GFAP-Cre Cre GFAP Mammalian Sofroniew 25997 LV-Cre pLKO.1 Cre and shRNA coexpression CMV Lentiviral Fuchs...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...

...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 [... Leong Ho 102970 pLKO.1P shFXN_1 FXN Friedreich ataxia Richard Possemato 102971 pLKO.1P shFXN_2 FXN Friedreich...HA-KIF5A KIF5A HA CMV ALS Rosalind Segal 167294 pLKO.1 puro_shTBK1_3185 TBK1 ALS Agnel Sfeir 169375 pET3d-SF-DCTN1...Cookson 13321 pGEX5X.1 PINK1 WT PINK1 GST tac Parkinson's Mark Cookson 13322 pGEX5X.1 PINK1 KD PINK1 GST... His, V5 EF-1 alpha Parkinson's Mark Cookson 25081 pDEST51-LRRK2-R1441C LRRK2 His, V5 EF-1 alpha Parkinson's... His, V5 EF-1 alpha Parkinson's Mark Cookson 25083 pDEST51-LRRK2-R1441H LRRK2 His, V5 EF-1 alpha Parkinson's...