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Species: Mus musculus
Genetic Insert: OKMS cassette
Vector Backbone Description: Vector Backbone:PB-TAC-Cas9; Vector Types:Mammalian Expression, CRISPR, piggyBac transposon; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30639212
Comments: Please Note: Addgene NGS is unable to fully resolve the CAG promoter sequence. Please refer to depositor's provided sequence for this section of the plasmid.
Proper citation: RRID:Addgene_120353 Copy
Species: Mus musculus
Genetic Insert: OK+9MS cassette
Vector Backbone Description: Vector Backbone:PB-TAC-Cas9; Vector Types:Mammalian Expression, CRISPR, piggyBac transposon; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30639212
Proper citation: RRID:Addgene_120354 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: Lipin-1
Vector Backbone Description: Vector Backbone:pRK5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30718857
Proper citation: RRID:Addgene_120277 Copy
Species: Mus musculus
Genetic Insert: Dicer
Vector Backbone Description: Vector Backbone:pEF1alpha.Blast; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_120541 Copy
Species: Mus musculus
Genetic Insert: ptch1
Vector Backbone Description: Vector Backbone:pVLAD6; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30415841
Proper citation: RRID:Addgene_120886 Copy
Species: Mus musculus
Genetic Insert: Foxo1
Vector Backbone Description: Backbone Size:4700; Vector Backbone:pCMV5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:16154098
Comments: Note that a BLAST search with Addgene's quality control sequence shows a K219R mutation. The Accili lab has explained that this is a common variant and they do not consider it a mutation at all. They have used the plasmid routinely without issue. The plasmid also contains an L619P polymorphism.
Proper citation: RRID:Addgene_12148 Copy
Species: Mus musculus
Genetic Insert: Foxo1
Vector Backbone Description: Backbone Size:4657; Vector Backbone:pCMV5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11696581
Comments: Plasmid contains K219R and L619P polymorphisms, which are not known to affect function.
Proper citation: RRID:Addgene_12142 Copy
Species: Mus musculus
Genetic Insert: ptch1
Vector Backbone Description: Vector Backbone:pcDNA-h; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30415841
Proper citation: RRID:Addgene_120903 Copy
Species: Mus musculus
Genetic Insert: p53
Vector Backbone Description: Backbone Size:5010; Vector Backbone:pLCR0; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:1761576
Comments: This plasmid, containing
the wild type murine p53 cDNA under the control of the previously
described BPV LCR-SV40 hybrid promoter (Spaholz et al., 1985),
was constructed as follows. Plasmid p407-1 (Spaholz et al., 1985) was
digested with EcoRI and HindIII, and the 5010-base pair fragment
containing the LCR-SV40 hybrid promoter was isolated and filled in
with Klenow.
After the ligation of BamHI linkers to the ends of the fragment, regenerating the terminal EcoRI site, the fragment was digested with
BamHI and ligated to the 2334-base pair BamHI fragment from the p53 cDNA expression vector pll-4 (Tan et al., 1986). This BamHI
fragment contained the entire wild type murine p53 coding region, 50 base pairs of 5'-untranslated sequence, 287 base pairs of 3'-untranslated
sequence, and a 3'-located SV40 mRNA splice site and polyA signal.
Proper citation: RRID:Addgene_12136 Copy
Species: Mus musculus
Genetic Insert: GBS(long)-GFP-insulator
Vector Backbone Description: Vector Backbone:pJC4-5; Vector Types:mouse transgenesis; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22265416
Proper citation: RRID:Addgene_121975 Copy
Species: Mus musculus
Genetic Insert: Myocardin
Vector Backbone Description: Backbone Marker:Clontech; Vector Backbone:pShuttle; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:19342595
Comments: Myocardin contains an A435T difference compared to the Myocd NCBI entry. The functional consequence of this difference is unknown.
Proper citation: RRID:Addgene_122122 Copy
Species: Mus musculus
Genetic Insert: MEKK3
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4700; Vector Backbone:pEYFP-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:14634666
Proper citation: RRID:Addgene_12189 Copy
Species: Mus musculus
Genetic Insert: MEKK1
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5500; Vector Backbone:pcDNA3.1/HisB; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:9528810
Comments: Please note that Addgene's sequencing results identified a V1467L mutation in the MEKK1 sequence when compared to GenBank ID AF117340.1. The depositing laboratory states that this plasmid functions exactly as intended and the mutation may represent a valid polymorphism.
Proper citation: RRID:Addgene_12181 Copy
Species: Mus musculus
Genetic Insert: MEKK2
Vector Backbone Description: Backbone Size:4600; Vector Backbone:pCMV5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:8621389
Proper citation: RRID:Addgene_12182 Copy
Species: Mus musculus
Genetic Insert: Hmga1a
Vector Backbone Description: Backbone Marker:Yukiko Gotoh, Addgene plasmid #122292; Vector Backbone:pCAG2IG; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22797695
Comments: Please note: Addgene NGS unable to fully resolve CAG promoter sequence. Please refer to the depositor's sequence for this section of the plasmid.
Proper citation: RRID:Addgene_122293 Copy
Species: Mus musculus
Genetic Insert: Mus musculus proprotein convertase subtilisin/kexin type 2 (Pcsk2)
Vector Backbone Description: Backbone Size:5428; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:30104250
Proper citation: RRID:Addgene_122673 Copy
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