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Species: Mus musculus
Genetic Insert: CCN1
Vector Backbone Description: Backbone Size:0; Vector Backbone:pAdTrace; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:16581771
Comments: pAdTrace is an RFP-based shuttle vector.
Proper citation: RRID:Addgene_12519 Copy
Species: Mus musculus
Genetic Insert: Wnt3A
Vector Backbone Description: Backbone Size:9220; Vector Backbone:pAdTrack-CMV; Vector Types:Mammalian Expression, Adenoviral; Bacterial Resistance:Kanamycin
Defining Citation: PMID:15496414
Proper citation: RRID:Addgene_12518 Copy
Species: Mus musculus
Genetic Insert: MCK promoter
Vector Backbone Description: Backbone Size:3000; Vector Backbone:pBluescriptKS; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:9844629
Comments: A 6.5 kb XhoI fragment containing 3 kb of the MCK promoter and enhancer 1, untranslated exon 1, 3 kb of intron 1 including the enhancer region E2, and the first 16 bp of exon 2 was subcloned into the XhoI site of pBluescriptKS to result in pBSMCK. A PolyA sequence was added downstream (see author's map, MCK-polyA-2). Please note that the restriction sites shown between the MCK promoter and polyA region in the author's map are NOT unique sites.
Proper citation: RRID:Addgene_12528 Copy
Species: Mus musculus
Genetic Insert: CCAAT/Enhancer-binding Protein beta
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5428; Vector Backbone:pcDNA3.1(-); Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Comments: The C/EBPbeta gene encodes 3 in-frame translational isoforms (LAP*, LAP, and LIP). LAP is the predominant activating isoform expressed in most cells, and this plasmid is designed to express the LAP isoform of the protein. The construct does not include the extra 21 codons at the N terminus that are present in LAP*. The complete insert (not just the ORF) includes a Kozak box sequence at the N terminus.
For more information on the C/EBPbeta isoforms, please see the original reference:
Descombes, P., and Schibler, U. A liver-enriched transcriptional activator protein, LAP, and a transcriptional inhibitory protein, LIP, are translated from the same mRNA. Cell 67:569-579 (1991).
Proper citation: RRID:Addgene_12557 Copy
Species: Mus musculus
Genetic Insert: Kir2.1
Vector Backbone Description: Vector Backbone:pCALNL; Vector Types:Mammalian Expression, Cre/Lox; Bacterial Resistance:Ampicillin
Defining Citation: PMID:37290446
Comments: Please visit https://www.biorxiv.org/content/10.1101/625616v2 for bioRxiv preprint.
Proper citation: RRID:Addgene_125578 Copy
Species: Mus musculus
Genetic Insert: Kir2.1
Vector Backbone Description: Vector Backbone:pCALNL; Vector Types:Mammalian Expression, Cre/Lox; Bacterial Resistance:Ampicillin
Defining Citation: PMID:37290446
Comments: Please visit https://www.biorxiv.org/content/10.1101/625616v2 for bioRxiv preprint.
Proper citation: RRID:Addgene_125579 Copy
Species: Mus musculus
Genetic Insert: CCAAT enhancer binding protein gamma
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5427; Vector Backbone:pcDNA3.1(-); Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11980905
Proper citation: RRID:Addgene_12560 Copy
Species: Mus musculus
Genetic Insert: shRNA targeting Sik3
Vector Backbone Description: Backbone Size:7900; Vector Backbone:LEPG #111160; Vector Types:Mammalian Expression, Retroviral, RNAi; Bacterial Resistance:Ampicillin
Defining Citation: PMID:31697837
Comments: Please visit https://www.biorxiv.org/content/10.1101/636969v1 for BioRxiv preprint
Proper citation: RRID:Addgene_125878 Copy
Species: Mus musculus
Genetic Insert: shRNA targeting Sik3
Vector Backbone Description: Backbone Size:7900; Vector Backbone:LEPG #111160; Vector Types:Mammalian Expression, Retroviral, RNAi; Bacterial Resistance:Ampicillin
Defining Citation: PMID:31697837
Comments: Please visit https://www.biorxiv.org/content/10.1101/636969v1 for BioRxiv preprint
Proper citation: RRID:Addgene_125855 Copy
Species: Mus musculus
Genetic Insert: Zif268 in pB1H1
Vector Backbone Description: Backbone Size:3527; Vector Backbone:pB1H1; Vector Types:Bacterial Expression; Bacterial Resistance:Chloramphenicol
Defining Citation: PMID:16041365
Proper citation: RRID:Addgene_12612 Copy
Species: Mus musculus
Genetic Insert: Type I procollagen α2 chain
Vector Backbone Description: Backbone Size:5400; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30287488
Comments: These mouse type I procollagen plasmids were designed for immortalized and primary murine osteoblasts and fibroblasts. Their expression in other cells might cause significant disruption of assembly, folding and trafficking of type I procollagen, resulting in artifacts. Even in osteoblasts and fibroblasts a fraction of transfected chains, particularly proα2, might be trafficked/degraded as monomers instead of being integrated into procollagen heterotrimers. In our experience, the best evidence of normal behavior of the transfected chains is the appearance of fluorescent extracellular collagen fibers ~ 12 h after transfection. Fluorescent molecules should be incorporated into fibers even when the transfected chains contain the N-propeptide cleavage site, because the cleavage is highly conformation sensitive and always incomplete.
We recommend following these guidelines when using the plasmids
1) Only mouse cells that have high expression of endogenous type I procollagen 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) Optimization of transfection efficiency is needed for all cell types.
4) Experiments beyond 24h after transfection are not recommended, to avoid excessive accumulation of aggregates and cell malfunction caused by increased procollagen synthesis.
5) Lack of extracellular fluorescent fibers 12-24 h after transfection indicates improper procollagen synthesis/trafficking or cellular malfunction.
6) 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_120160 Copy
Species: Mus musculus
Genetic Insert: kinesin family member 5A
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4727; Vector Backbone:pmCherry-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:31100061
Proper citation: RRID:Addgene_120163 Copy
Species: Mus musculus
Genetic Insert: kinesin family member 1A
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4727; Vector Backbone:pmCherry-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:31100061
Proper citation: RRID:Addgene_120165 Copy
Species: Mus musculus
Genetic Insert: BICD cargo adaptor 2
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4713; Vector Backbone:pmCherry-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:31100061
Proper citation: RRID:Addgene_120168 Copy
Species: Mus musculus
Genetic Insert: Vrk2
Vector Backbone Description: Vector Backbone:pLV-EF1a-IRES-Puro; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:31142202
Proper citation: RRID:Addgene_120240 Copy
Species: Mus musculus
Genetic Insert: Sec61B
Vector Backbone Description: Vector Backbone:pLV-EF1a-IRES-Puro; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:31142202
Proper citation: RRID:Addgene_120244 Copy
Species: Mus musculus
Genetic Insert: Lem2
Vector Backbone Description: Vector Backbone:pLV-EF1a-IRES-Puro; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:31142202
Proper citation: RRID:Addgene_120245 Copy
Species: Mus musculus
Genetic Insert: Mfsd10
Vector Backbone Description: Vector Backbone:pLV-EF1a-IRES-Puro; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:31142202
Proper citation: RRID:Addgene_120238 Copy
Species: Mus musculus
Genetic Insert: Tmem128
Vector Backbone Description: Vector Backbone:pLV-EF1a-IRES-Puro; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_120239 Copy
Species: Mus musculus
Genetic Insert: Arl6ip6
Vector Backbone Description: Vector Backbone:pLV-EF1a-IRES-Puro; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:31142202
Proper citation: RRID:Addgene_120235 Copy
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