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Species: Homo sapiens
Genetic Insert: EGFR
Vector Backbone Description: Backbone Size:5169; Vector Backbone:pBABEpuro; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17495523
Proper citation: RRID:Addgene_32069 Copy
Species: Homo sapiens
Genetic Insert: EGFR
Vector Backbone Description: Backbone Size:5169; Vector Backbone:pBABEpuro; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17495523
Proper citation: RRID:Addgene_32063 Copy
Species: Homo sapiens
Genetic Insert: EGFR
Vector Backbone Description: Backbone Size:5169; Vector Backbone:pBABEpuro; Vector Types:Mammalian Expression, Retroviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:17495523
Proper citation: RRID:Addgene_32064 Copy
Species: Homo sapiens
Genetic Insert: HDAC5
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:37000; Vector Backbone:pAd CMV/V5 DEST; Vector Types:Mammalian Expression, Adenoviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21565617
Proper citation: RRID:Addgene_32218 Copy
Species: Homo sapiens
Genetic Insert: HDAC5
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:37000; Vector Backbone:pAd CMV/V5 DEST; Vector Types:Mammalian Expression, Adenoviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21565617
Proper citation: RRID:Addgene_32208 Copy
Species: Homo sapiens
Genetic Insert: GSK3B
Vector Backbone Description: Backbone Marker:Sabatini Lab; Backbone Size:7000; Vector Backbone:pLKO.1; Vector Types:Mammalian Expression, Lentiviral, RNAi; Bacterial Resistance:Ampicillin
Defining Citation: PMID:19258413
Proper citation: RRID:Addgene_32496 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-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: Synapsin I promoter
Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pDONR221 P1-P5r; Vector Types:Gateway Cloning; Bacterial Resistance:Kanamycin
Defining Citation: PMID:21772812
Comments: Gene Therapy 14, 872-882 (June 2007) doi:10.1038/sj.gt.3302924
Efficient gene transduction of neurons by lentivirus with enhanced neuron-specific promoters
H Hioki, H Kameda, H Nakamura, T Okunomiya, K Ohira, K Nakamura, M Kuroda, T Furuta and T Kaneko
Proper citation: RRID:Addgene_32582 Copy
Species: Homo sapiens
Genetic Insert: CCND1 promoter
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:4818; Vector Backbone:pGL3Basic; Vector Types:Luciferase; Bacterial Resistance:Ampicillin
Defining Citation: PMID:10201372
Proper citation: RRID:Addgene_32731 Copy
Species: Homo sapiens
Genetic Insert: CCND1 promoter
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:4818; Vector Backbone:pGL3Basic; Vector Types:Luciferase; Bacterial Resistance:Ampicillin
Defining Citation: PMID:10201372
Proper citation: RRID:Addgene_32734 Copy
Species: Homo sapiens
Genetic Insert: CCND1 promoter
Vector Backbone Description: Backbone Marker:Promega; Backbone Size:4818; Vector Backbone:pGL3Basic; Vector Types:Luciferase; Bacterial Resistance:Ampicillin
Defining Citation: PMID:10201372
Comments: There is a 1bp mismatch in the sequence that does not affect plasmid function.
Proper citation: RRID:Addgene_32737 Copy
Species: Homo sapiens
Genetic Insert: MLH1
Vector Backbone Description: Backbone Marker:Structural Genomics Consortium (Addgene plasmid 26096); Backbone Size:7325; Vector Backbone:pET28-MHL; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: PDB: 3RBN. http://www.thesgc.org/structures/3RBN/
Proper citation: RRID:Addgene_32859 Copy
Species: Homo sapiens
Genetic Insert: OCLN
Vector Backbone Description: Backbone Marker:Gungxia Gao; Vector Backbone:pTrip; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20822789
Proper citation: RRID:Addgene_32794 Copy
Species: Homo sapiens
Genetic Insert: OCLN
Vector Backbone Description: Backbone Marker:Gungxia Gao; Vector Backbone:pTrip; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20822789
Proper citation: RRID:Addgene_32792 Copy
Species: Homo sapiens
Genetic Insert: OCLN
Vector Backbone Description: Backbone Marker:Gungxia Gao; Vector Backbone:pTrip; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20822789
Proper citation: RRID:Addgene_32797 Copy
Species: Homo sapiens
Genetic Insert: AMOT p80
Vector Backbone Description: Vector Backbone:pHRCTS-CMV-WPRE-GFP; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:21205866
Proper citation: RRID:Addgene_32830 Copy
Species: Homo sapiens
Genetic Insert: OCLN
Vector Backbone Description: Backbone Marker:Gungxia Gao; Vector Backbone:pTrip; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:20822789
Proper citation: RRID:Addgene_32796 Copy
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