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Species: Rattus norvegicus
Genetic Insert: Constitutively active Rheb (S16H)
Vector Backbone Description: Backbone Marker:Addgene plasmid #32519; Vector Backbone:pHAGE-CMV-Rheb-IRES-eGFP-W; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20062052
Comments: The constitutively activating S16H mutation was introduced to pHAGE-CMV-Rheb-IRES-eGFP-W (Addgene plasmid #32519) by site directed mutagenesis.
The original pHAGE-CMV-IRES lentiviral vector is from Dr Richard C. Mulligan at Harvard (Mostoslavsky G et al., 2005).
Proper citation: RRID:Addgene_32520 Copy
Species: Homo sapiens
Genetic Insert: PSAML141F,Y115F-GlyR
Vector Backbone Description: Backbone Size:5100; Vector Backbone:AAV2; Vector Types:AAV, Cre/Lox, Adeno-associated virus, FLEX switch; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21885782
Comments: Comments:
These rAAV vectors are prone to recombination. This is a well known issue with these rAAV vectors and is due to the inverted terminal repeats (ITRs) required for rAAV production. To minimize recombination, we propagate these plasmids in NEB Stable cells. Also, to minimize recombination, cells should be cultured at 30 C.
Note that these cultures will grow slowly (20 h for minipreps). Better yields and culture times are obtained with 2xYT as the media. This is strongly recommended.
Because recombination may still happen, we do a panel of restriction digestions to assess whether the ITRs are in tact. Separate digestions with Sma1 and Msc1 should be performed. The expected patterns can be calculated from the attached sequence.
In Magnus et al, the PSAMY115F,L141F-GlyR channel was emphasized for silencing because this channel has the lowest acetylcholine responsiveness. However, PSAML141F-GlyR constructs, lacking the Y115F mutation, already have low ACh potency. The Y115F mutation also slightly increases the EC50 for the activator ligand, PSEM89S. Since PSAML141F-GlyR constructs already have very low acetylcholine potency, in many cases it may not be worth the slight reduction in PSEM potency. We have made both constructs available for researchers who may find that they have different requirements with respect to acetylcholine responsiveness.
Proper citation: RRID:Addgene_32481 Copy
Species: Homo sapiens
Genetic Insert: Protein tyrosine phosphatase receptor-type delta G1707R
Vector Backbone Description: Backbone Marker:System Biosciences ; Backbone Size:6606; Vector Backbone:pCDF1-MCS2-EF1-Puro; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19074898
Proper citation: RRID:Addgene_32526 Copy
Species: Homo sapiens
Genetic Insert: Protein tyrosine phosphatase receptor-type delta P1690F
Vector Backbone Description: Backbone Marker:System Biosciences ; Backbone Size:6606; Vector Backbone:pCDF1-MCS2-EF1-Puro; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19074898
Proper citation: RRID:Addgene_32525 Copy
Species: Homo sapiens
Genetic Insert: PSAMQ79G,Q139G-5HT3 HC
Vector Backbone Description: Backbone Size:7200; Vector Backbone:pCAGGS- IRES-GFP; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21885782
Proper citation: RRID:Addgene_32474 Copy
Species: Homo sapiens
Genetic Insert: PSAMQ79G,Q139G-5HT3HC
Vector Backbone Description: Backbone Size:5100; Vector Backbone:AAV2; Vector Types:AAV, Cre/Lox, Adeno-associated virus, FLEX switch; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21885782
Comments: These rAAV vectors are prone to recombination. This is a well known issue with these rAAV vectors and is due to the inverted terminal repeats (ITRs) required for rAAV production. To minimize recombination, we propagate these plasmids in NEB Stable cells. Also, to minimize recombination, cells should be cultured at 30 C.
Note that these cultures will grow slowly (20 h for minipreps). Better yields and culture times are obtained with 2xYT as the media. This is strongly recommended.
Because recombination may still happen, we do a panel of restriction digestions to assess whether the ITRs are in tact. Separate digestions with Sma1 and Msc1 should be performed. The expected patterns can be calculated from the attached sequence.
Proper citation: RRID:Addgene_32475 Copy
Species: Homo sapiens
Genetic Insert: PSAM Y115F,L141F-5HT3 HC
Vector Backbone Description: Backbone Size:7200; Vector Backbone:pCAGGS- IRES-GFP; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21885782
Proper citation: RRID:Addgene_32476 Copy
Vector Backbone Description: Backbone Size:12000; Vector Backbone:pCAMBIA1301; Vector Types:Plant Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:20388670
Proper citation: RRID:Addgene_32470 Copy
Vector Backbone Description: Backbone Size:12000; Vector Backbone:pCAMBIA1301; Vector Types:Plant Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:20388670
Proper citation: RRID:Addgene_32471 Copy
Species: Mus musculus
Genetic Insert: Pax7d
Vector Backbone Description: Backbone Marker:Addgene plasmid #32516; Backbone Size:7900; Vector Backbone:pOZ-FH-C-Puro; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19458195
Proper citation: RRID:Addgene_32518 Copy
Species: synthetic construct
Genetic Insert: GEX-GECO1.0
Vector Backbone Description: Backbone Size:4200; Vector Backbone:Customized Vector; Vector Types:Bacterial Expression, Lab constructed; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21903779
Comments: The vector is modified from pBAD/ His B (Invitrogen).
Proper citation: RRID:Addgene_32464 Copy
Genetic Insert: G-GECO1.1
Vector Backbone Description: Backbone Size:4100; Vector Backbone:Customized Vector; Vector Types:Bacterial Expression, Lab constructed; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21903779
Comments: The vector is modified from pBAD/ His B (Invitrogen).
Proper citation: RRID:Addgene_32467 Copy
Genetic Insert: G-GECO1.2
Vector Backbone Description: Backbone Size:4200; Vector Backbone:Customized Vector; Vector Types:Bacterial Expression, Lab constructed; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21903779
Comments: The vector is modified from pBAD/ His B (Invitrogen).
Proper citation: RRID:Addgene_32468 Copy
Species: synthetic construct
Genetic Insert: R-GECO1.0
Vector Backbone Description: Backbone Size:4200; Vector Backbone:Customized Vector; Vector Types:Bacterial Expression, Lab constructed; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21903779
Comments: The vector is modified from pBAD/ His B (Invitrogen).
Proper citation: RRID:Addgene_32465 Copy
Species: Homo sapiens
Genetic Insert: NRBP
Vector Backbone Description: Backbone Marker:Agilent; Backbone Size:4100; Vector Backbone:pSG5; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11956649
Proper citation: RRID:Addgene_32504 Copy
Species: Homo sapiens
Genetic Insert: CIB1
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5400; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:11756406
Proper citation: RRID:Addgene_32505 Copy
Genetic Insert: BCR
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:5400; Vector Backbone:pcDNA3.1; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17116687
Proper citation: RRID:Addgene_32507 Copy
Species: Homo sapiens
Genetic Insert: KLF4
Vector Backbone Description: Backbone Size:4600; Vector Backbone:pMXs; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:18035408
Comments: The sequence of primer pMX-S1811 is GAC GGC ATC GCA GCT TGG ATA CAC.
Proper citation: RRID:Addgene_17219 Copy
Species: Homo sapiens
Genetic Insert: c-MYC
Vector Backbone Description: Backbone Size:4600; Vector Backbone:pMXs; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:18035408
Comments: The sequence of primer pMX-S1811 is GAC GGC ATC GCA GCT TGG ATA CAC.
Proper citation: RRID:Addgene_17220 Copy
Species: Homo sapiens
Genetic Insert: Upstream region of human REX1
Vector Backbone Description: Backbone Size:4842; Vector Backbone:pGV-BM2; Vector Types:Mammalian Expression, Luciferase; Bacterial Resistance:Ampicillin
Defining Citation: PMID:18035408
Proper citation: RRID:Addgene_17222 Copy
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