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http://www.addgene.org/140100

Species: Homo sapiens
Genetic Insert: gRNA1+scaffold+H1promoter+gRNA2
Vector Backbone Description: Vector Backbone:pDECKO_mCherry; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:35619054

Proper citation: RRID:Addgene_140100 Copy   

•   Source: Addgene

http://www.addgene.org/140188

Species: Synthetic
Genetic Insert: Salsa6f
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:3923; Vector Backbone:pEGFP-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:29239725
Comments: The fusion protein in this plasmid is the fluorescent genetically encoded calcium indicator Salsa6f, which is a fusion of the dimeric red fluorescent protein tdTomato, the epitope tag V5, and the green fluorescent genetically encoded calcium indicator GCaMP6f, in order from N- to C-termini. The presence of tdTomato enables visualization of transfected cells in the absence of Ca2+ signaling and concentration-independent, ratiometric readout of Ca2+ signals. Salsa6f has been introduced into transgenic mice, where its expression is under Cre recombinase control (see PMID: 29239725).

Proper citation: RRID:Addgene_140188 Copy   

•   Source: Addgene

http://www.addgene.org/140101

Species: Homo sapiens
Genetic Insert: gRNA1+scaffold+H1promoter+gRNA2
Vector Backbone Description: Vector Backbone:pDECKO_mCherry; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:35619054

Proper citation: RRID:Addgene_140101 Copy   

•   Source: Addgene

http://www.addgene.org/140102

Species: Homo sapiens
Genetic Insert: gRNA1+scaffold+H1promoter+gRNA2
Vector Backbone Description: Vector Backbone:pDECKO_mCherry; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:35619054

Proper citation: RRID:Addgene_140102 Copy   

•   Source: Addgene

http://www.addgene.org/140103

Species: Homo sapiens
Genetic Insert: gRNA1+scaffold+H1promoter+gRNA2
Vector Backbone Description: Vector Backbone:pDECKO_mCherry; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:35619054

Proper citation: RRID:Addgene_140103 Copy   

•   Source: Addgene

http://www.addgene.org/140186

Vector Backbone Description: Backbone Size:4483; Vector Backbone:pEU3-NII-GLICNot-C-BIOT; Vector Types:expression vector for a cell-free system; Bacterial Resistance:Ampicillin
Defining Citation: PMID:32169064

Proper citation: RRID:Addgene_140186 Copy   

•   Source: Addgene

http://www.addgene.org/140187

Vector Backbone Description: Backbone Size:4456; Vector Backbone:pEU3-NII-GLICNot-C-His; Vector Types:expression vector for a cell-free system; Bacterial Resistance:Ampicillin
Defining Citation: PMID:32169064

Proper citation: RRID:Addgene_140187 Copy   

•   Source: Addgene

http://www.addgene.org/140182

Vector Backbone Description: Backbone Size:4444; Vector Backbone:pEU3-NII-GLICNot; Vector Types:expression vector for a cell-free system; Bacterial Resistance:Ampicillin
Defining Citation: PMID:32169064

Proper citation: RRID:Addgene_140182 Copy   

•   Source: Addgene

http://www.addgene.org/140183

Vector Backbone Description: Backbone Size:3865; Vector Backbone:pEU3-NII-HxHLICNot; Vector Types:expression vector for a cell-free system; Bacterial Resistance:Ampicillin
Defining Citation: PMID:32169064

Proper citation: RRID:Addgene_140183 Copy   

•   Source: Addgene

http://www.addgene.org/140108

Species: Homo sapiens
Genetic Insert: gRNA1+scaffold+H1promoter+gRNA2
Vector Backbone Description: Vector Backbone:pDECKO_mCherry; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:35619054

Proper citation: RRID:Addgene_140108 Copy   

•   Source: Addgene

http://www.addgene.org/140104

Species: Homo sapiens
Genetic Insert: gRNA1+scaffold+H1promoter+gRNA2
Vector Backbone Description: Vector Backbone:pDECKO_mCherry; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:35619054

Proper citation: RRID:Addgene_140104 Copy   

•   Source: Addgene

http://www.addgene.org/140106

Species: Homo sapiens
Genetic Insert: gRNA1+scaffold+H1promoter+gRNA2
Vector Backbone Description: Vector Backbone:pDECKO_mCherry; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:35619054

Proper citation: RRID:Addgene_140106 Copy   

•   Source: Addgene

http://www.addgene.org/140107

Species: Homo sapiens
Genetic Insert: gRNA1+scaffold+H1promoter+gRNA2
Vector Backbone Description: Vector Backbone:pDECKO_mCherry; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:35619054

Proper citation: RRID:Addgene_140107 Copy   

•   Source: Addgene

http://www.addgene.org/140111

Species: Homo sapiens
Genetic Insert: Type I procollagen α1
Vector Backbone Description: Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: These human type I procollagen plasmids were designed and validated for human osteoblasts and fibroblasts similar to their previously published/shared murine counterparts e.g. eGFP-Proα1 (Plasmid #119843) https://www.addgene.org/Sergey_Leikin/. Their expression in other cells might disrupt assembly, folding and trafficking of type I procollagen, resulting in artifacts. Even in human osteoblasts and fibroblasts, overexpression of exogenous proα2 might cause intracellular trafficking and degradation of some transfected chains as monomers without integration into normal heterotrimeric (proα1)2proα2 procollagen molecules,. Overexpression of exogenous proα1 might result in excessive formation of homotrimeric (proα1)3 molecules, some of which might contain fluorescent tags on two or even all three proα1 chains, potentially causing severe disruptions in cellular function. In our experience with the murine plasmids expressed in primary osteoblasts or the MC3T3 osteoblastic cell line, the best evidence of normal behavior of the transfected chains is the appearance of fluorescent extracellular collagen fibers ~ 12-24 h after transfection. These were harder to observe in human Saos2 osteoblast cell line transfected with the human type I procollagen plasmids (likely due to slower fibrillogenesis and more complete cleavage of the fluorescently-tagged N-propeptide prior to collagen fibril formation). We strongly recommend following these guidelines when using the plasmids 1) Only human cells that have high expression of endogenous type I procollagen and that express fewer transfected than endogenous chains are suitable for studies of physiologically-relevant processes with these constructs. 2) At least 100 μM ascorbic acid at and after transfection is required to ensure proper procollagen folding (for some cells, preincubation with ascorbic acid before transfection might be needed). 3) Stable transfection causes excessive accumulation of tagged chains in the ER over time, disruption of ER homeostasis, and disruptions and changes in cellular function. Therefore, low to moderate transient transfection is recommended for studies of physiologically relevant processes. Optimization of transfection efficiency is needed for all cell types. 4) Experiments beyond 24h after transfection are not recommended, to avoid the excessive accumulation of tagged chains in the ER and cell malfunction caused by increased procollagen synthesis. 5) Excessive expression of transfected chains might cause rapid accumulation of large procollagen aggregates in the ER, resulting in cellular malfunction and death. Contact information: Shakib Omari ([email protected]), Sergey Leikin ([email protected]).

Proper citation: RRID:Addgene_140111 Copy   

•   Source: Addgene

http://www.addgene.org/140079

Species: Mus musculus
Genetic Insert: anti-GIT2 (Human) recombinant mouse monoclonal antibody
Vector Backbone Description: Backbone Marker:Yves Durocher, NRC; Backbone Size:5925; Vector Backbone:P1316-IgG2a; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30667360
Comments: RRID: AB_2750816. Isotype: IgG2a. Additional Species: human, mouse, rat. A portion of this plasmid was derived from a plasmid, pTT3, which was obtained from Yves Durocher, National Research Council of Canada- Biotechnology Research Institute.

Proper citation: RRID:Addgene_140079 Copy   

•   Source: Addgene

http://www.addgene.org/140112

Species: Homo sapiens
Genetic Insert: Type I procollagen α1
Vector Backbone Description: Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: These human type I procollagen plasmids were designed and validated for human osteoblasts and fibroblasts similar to their previously published/shared murine counterparts e.g. eGFP-Proα1 (Plasmid #119843) https://www.addgene.org/Sergey_Leikin/. Their expression in other cells might disrupt assembly, folding and trafficking of type I procollagen, resulting in artifacts. Even in human osteoblasts and fibroblasts, overexpression of exogenous proα2 might cause intracellular trafficking and degradation of some transfected chains as monomers without integration into normal heterotrimeric (proα1)2proα2 procollagen molecules,. Overexpression of exogenous proα1 might result in excessive formation of homotrimeric (proα1)3 molecules, some of which might contain fluorescent tags on two or even all three proα1 chains, potentially causing severe disruptions in cellular function. In our experience with the murine plasmids expressed in primary osteoblasts or the MC3T3 osteoblastic cell line, the best evidence of normal behavior of the transfected chains is the appearance of fluorescent extracellular collagen fibers ~ 12-24 h after transfection. These were harder to observe in human Saos2 osteoblast cell line transfected with the human type I procollagen plasmids (likely due to slower fibrillogenesis and more complete cleavage of the fluorescently-tagged N-propeptide prior to collagen fibril formation). We strongly recommend following these guidelines when using the plasmids 1) Only human cells that have high expression of endogenous type I procollagen and that express fewer transfected than endogenous chains are suitable for studies of physiologically-relevant processes with these constructs. 2) At least 100 μM ascorbic acid at and after transfection is required to ensure proper procollagen folding (for some cells, preincubation with ascorbic acid before transfection might be needed). 3) Stable transfection causes excessive accumulation of tagged chains in the ER over time, disruption of ER homeostasis, and disruptions and changes in cellular function. Therefore, low to moderate transient transfection is recommended for studies of physiologically relevant processes. Optimization of transfection efficiency is needed for all cell types. 4) Experiments beyond 24h after transfection are not recommended, to avoid the excessive accumulation of tagged chains in the ER and cell malfunction caused by increased procollagen synthesis. 5) Excessive expression of transfected chains might cause rapid accumulation of large procollagen aggregates in the ER, resulting in cellular malfunction and death. Contact information: Shakib Omari ([email protected]), Sergey Leikin ([email protected]).

Proper citation: RRID:Addgene_140112 Copy   

•   Source: Addgene

http://www.addgene.org/140196

Species: Synthetic
Genetic Insert: dSpCas9-VPR
Vector Backbone Description: Backbone Size:3033; Vector Backbone:pGem; Vector Types:CRISPR, Synthetic Biology, Fungal genome integration.; Bacterial Resistance:Ampicillin
Defining Citation: PMID:32526136
Comments: To be linerised with NotI for homologous recombination in Aspergillus nidulans nkuA deficient strains, selecting with glufosinate/Basta. It can also be used for random genome integration in other fungal strains.

Proper citation: RRID:Addgene_140196 Copy   

•   Source: Addgene

http://www.addgene.org/140197

Species: Synthetic
Genetic Insert: dLbCas12a-VPR
Vector Backbone Description: Backbone Marker:DOI: 10.1039/C8SC02870B; Backbone Size:14777; Vector Backbone:pYFAC-riboB; Vector Types:CRISPR, Synthetic Biology, Expression in Aspergillus nidulans and related filamentous fungi.; Bacterial Resistance:Ampicillin and Kanamycin
Defining Citation: PMID:32526136

Proper citation: RRID:Addgene_140197 Copy   

•   Source: Addgene

http://www.addgene.org/140089

Species: Other
Genetic Insert: DENV2A-GFP
Vector Backbone Description: Backbone Marker:Invitrogen modified by Ie-Ming Shih; Backbone Size:7057; Vector Backbone:pLenti-puro (Addgene Plasmid #39481); Vector Types:Mammalian Expression, Lentiviral, Low expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:31919100
Comments: The DENV2A-GFP reporter is not suitable for transient transfection as the reporter has a basal background that makes it sensitive to the levels of expression: If the expression is too high the background will mask the signal produced by the reporter - If the expression is too low you won’t detect enough signal over the background. Our recommendation is to use the pLenti-DENV2A-GFP-puro construct to pack lentiviral particles with the psPAX2 (Addgene #12260) and pMD2.G (Addgene #12259) plasmids, generate stable cell lines and sort cell subpopulations with different levels of the reporter's basal background before testing them. The best reporter cells will be those with the highest signal-to-noise ratio upon DENV-2 infection.

Proper citation: RRID:Addgene_140089 Copy   

•   Source: Addgene

http://www.addgene.org/140085

Species: Lactate oxidase from A. viridans and Catalase from E. coli
Genetic Insert: A fusion of lactate oxidase from A. viridans and catalase from E. coli
Vector Backbone Description: Backbone Size:8838; Vector Backbone:pET30a; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:31932725

Proper citation: RRID:Addgene_140085 Copy   

•   Source: Addgene