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Species: Mus musculus
Genetic Insert: ret
Vector Backbone Description: Vector Backbone:pBS; Vector Types:in situ hybridization probe; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20064391
Comments: This plasmid can be used to generate an in situ hybridization antisense probe for mouse C-ret. Linearize the plasmid with NotI and use T7 to transcribe the ~1kb probe.
Proper citation: RRID:Addgene_119900 Copy
Species: Mus musculus
Genetic Insert: Type I procollagen α1 chain
Vector Backbone Description: Backbone Size:5300; 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_119843 Copy
Species: Mus musculus
Genetic Insert: Type I procollagen α1 chain
Vector Backbone Description: Backbone Size:5300; 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_119844 Copy
Species: Mus musculus
Genetic Insert: Type I procollagen α1 chain
Vector Backbone Description: Backbone Size:5300; 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_119842 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_119840 Copy
Species: Mus musculus
Genetic Insert: Sik3
Vector Backbone Description: Backbone Size:7210; Vector Backbone:LentiV_Neo; Vector Types:Mammalian Expression, Lentiviral; 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_119973 Copy
Species: Mus musculus
Genetic Insert: Sik3
Vector Backbone Description: Backbone Size:7210; Vector Backbone:LentiV_Neo; Vector Types:Mammalian Expression, Lentiviral; 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_119972 Copy
Species: Mus musculus
Genetic Insert: LIN28A
Vector Backbone Description: Vector Backbone:pCCL; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:27295554
Comments: This plasmid was generated by Mario Leonardo Squadrito.
Proper citation: RRID:Addgene_117057 Copy
Species: Mus musculus
Genetic Insert: Tet3
Vector Backbone Description: Backbone Size:3000; Vector Backbone:pDT; Vector Types:Mouse Targeting; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_117146 Copy
Species: Mus musculus
Genetic Insert: Nkx2-5 CDS
Vector Backbone Description: Backbone Size:9105; Vector Backbone:pKLV2-U6(BsmBI)-EF1a-BFP-Puro-Cas9(FZ); Vector Types:Lentiviral, CRISPR; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30932271
Proper citation: RRID:Addgene_117135 Copy
Species: Mus musculus
Genetic Insert: Nkx2-5 CDS
Vector Backbone Description: Backbone Size:9601; Vector Backbone:pKLV2-U6(BsmBI)-EF1a-BFP-BSD; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30932271
Proper citation: RRID:Addgene_117137 Copy
Species: Mus musculus
Genetic Insert: partial sequence of mannosidase II
Vector Backbone Description: Backbone Marker:Clontech; Vector Backbone:pIRESneo3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30587510
Proper citation: RRID:Addgene_117274 Copy
Species: Mus musculus
Genetic Insert: Egr-1
Vector Backbone Description: Backbone Size:5400; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:15225550
Proper citation: RRID:Addgene_11729 Copy
Species: Mus musculus
Genetic Insert: Sox9
Vector Backbone Description: Backbone Size:9063; Vector Backbone:Tet-O-FUW; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30127505
Proper citation: RRID:Addgene_117269 Copy
Species: Mus musculus
Genetic Insert: Nfib
Vector Backbone Description: Backbone Size:10013; Vector Backbone:Tet-O-FUW; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30127505
Proper citation: RRID:Addgene_117271 Copy
Species: Mus musculus
Genetic Insert: ELMOD3
Vector Backbone Description: Vector Backbone:eneArt vector; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_117443 Copy
Species: Mus musculus
Genetic Insert: Activin receptor IIA
Vector Backbone Description: Backbone Size:4700; Vector Backbone:pCMV5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:9748228
Proper citation: RRID:Addgene_11754 Copy
Species: Mus musculus
Genetic Insert: ALK6
Vector Backbone Description: Backbone Size:4700; Vector Backbone:pCMV5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:9748228
Proper citation: RRID:Addgene_11769 Copy
Species: Mus musculus
Genetic Insert: Sod1
Vector Backbone Description: Backbone Marker:Novagen; Vector Backbone:pET28a(+); Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Proper citation: RRID:Addgene_117710 Copy
Species: Mus musculus
Genetic Insert: Sod1
Vector Backbone Description: Backbone Marker:Novagen; Vector Backbone:pET28a(+); Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Proper citation: RRID:Addgene_117701 Copy
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