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Species: Homo sapiens
Genetic Insert: Wnt reporter sequences
Vector Backbone Description: Backbone Marker:addgene; Vector Backbone:pSicoR PGK Puro; Vector Types:Mammalian Expression, Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22177620
Comments: This lentiviral Wnt reporter has 7 repeats of Tcf/Lef binding sites, which was cloned by Xiaojun (Lance) Lian.
Methods for Plasmid construction, lentiviral production and infection of hESCs:
The 7 TCF/LEF binding site sequences were obtained by direct PCR of plasmid M50 (Addgene plasmid 12456) and the GreenFire (GF) sequences were obtained by direct PCR of the pGreenFire1-mCMV Plasmid (System Bioscience TR010PA-1). These two sequences were cloned into pSicoR PGK puro (Addgene plasmid 11586) digested by Xba I (NEB) and Xho I (NEB). The constructed Wnt reporter plasmid was named 7TGFP and verified by sequencing. This 7TGFP vector was cotransfected with the helper plasmids psPAX2 and pMD2.G (Addgene plasmids 12260 and 12259) into HEK-293TN cells (System Biosciences) for virus production. Virus-containing medium was collected at 48 and 72 hours after transfection and used for infection of hESCs in the presence of 6μg/mL polybrene (Sigma). Transduced cells were cultured in mTeSR1 on Matrigel for three days and then clonally isolated in mTeSR1 with 1 μg/mL puromycin.
Proper citation: RRID:Addgene_40588 Copy
Species: Homo sapiens
Genetic Insert: TCF7
Vector Backbone Description: Backbone Size:5400; Vector Backbone:pcDNA3; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22949634
Proper citation: RRID:Addgene_40620 Copy
Species: Homo sapiens
Genetic Insert: PA1
Vector Backbone Description: Backbone Marker:System Biosciences (SBI); Backbone Size:8295; Vector Backbone:pCDH-EF1-MCS-IRES-Neo; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22949634
Proper citation: RRID:Addgene_40618 Copy
Species: Homo sapiens
Genetic Insert: RPRD1B
Vector Backbone Description: Backbone Marker:SGC; Vector Backbone:pET15-MHL; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Comments: http://www.thesgc.org/structures/4FLA/
Proper citation: RRID:Addgene_40739 Copy
Species: Homo sapiens
Genetic Insert: L3MBTL3
Vector Backbone Description: Backbone Marker:SGC; Vector Backbone:pNIC-CH; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: http://www.thesgc.org/structures/4FL6/
Proper citation: RRID:Addgene_40738 Copy
Species: Homo sapiens
Genetic Insert: LRRK2
Vector Backbone Description: Backbone Marker:Emerald; Vector Backbone:pEMB108; Vector Types:Baculovirus; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_40567 Copy
Species: Homo sapiens
Genetic Insert: LRRK2
Vector Backbone Description: Backbone Marker:Emerald; Vector Backbone:pEMB52; Vector Types:Baculovirus; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_40565 Copy
Species: Homo sapiens
Genetic Insert: LRRK2
Vector Backbone Description: Backbone Marker:Emerald; Vector Backbone:pEMB12; Vector Types:Baculovirus; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_40564 Copy
Species: Homo sapiens
Genetic Insert: DVL1
Vector Backbone Description: Backbone Marker:Emerald; Vector Backbone:pEMB36; Vector Types:Baculovirus; Bacterial Resistance:Ampicillin
Comments: The insert has been codon optimized.
Proper citation: RRID:Addgene_40557 Copy
Species: Homo sapiens
Genetic Insert: LRRK2
Vector Backbone Description: Backbone Marker:Emerald; Vector Backbone:pEMB12; Vector Types:Baculovirus; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_40553 Copy
Species: Homo sapiens
Genetic Insert: DVL1
Vector Backbone Description: Backbone Marker:Emerald; Vector Backbone:pEMB52; Vector Types:Baculovirus; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_40542 Copy
Species: Homo sapiens
Genetic Insert: YWHAG
Vector Backbone Description: Backbone Marker:Emerald; Vector Backbone:pEMB52; Vector Types:Baculovirus; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_40541 Copy
Species: Homo sapiens
Genetic Insert: sh-hSOX9-1
Vector Backbone Description: Backbone Marker:Weinberg lab; Backbone Size:7032; Vector Backbone:pLKO.1 puro (Addgene plasmid 8453); Vector Types:Mammalian Expression, Lentiviral, RNAi; Bacterial Resistance:Ampicillin
Defining Citation: PMID:22385965
Proper citation: RRID:Addgene_40644 Copy
Species: Homo sapiens
Genetic Insert: SNCA
Vector Backbone Description: Backbone Marker:Roche; Backbone Size:5450; Vector Backbone:pHM6; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:10698681
Proper citation: RRID:Addgene_40825 Copy
Species: Homo sapiens
Genetic Insert: SNCA
Vector Backbone Description: Backbone Marker:Roche; Backbone Size:5450; Vector Backbone:pHM6; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:10698681
Proper citation: RRID:Addgene_40824 Copy
Species: Homo sapiens
Genetic Insert: SNCA
Vector Backbone Description: Backbone Marker:Clontech; Backbone Size:4700; Vector Backbone:pEGFP-C1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Defining Citation: PMID:10698681
Comments: Please note that EcoRI is not a unique site. The alpha-synuclein insert contains an EcoRI site in the coding sequence.
Proper citation: RRID:Addgene_40822 Copy
Species: Homo sapiens
Genetic Insert: XIAP
Vector Backbone Description: Backbone Marker:Baltimore Lab; Backbone Size:5353; Vector Backbone:pEBB; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:14701799
Comments: Site-directed mutagenesis was used to to introduce a stop codon in pEBB XIAP at AA 449 (AAG to TGA).
(#721)
Proper citation: RRID:Addgene_40828 Copy
Species: Homo sapiens
Genetic Insert: MAP2K1
Vector Backbone Description: Backbone Marker:Invitrogen; Backbone Size:10200; Vector Backbone:pCEP4; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:8052857
Proper citation: RRID:Addgene_40811 Copy
Species: Homo sapiens
Genetic Insert: Braf
Vector Backbone Description: Backbone Size:5600; Vector Backbone:pcDNA3.1-Hygro; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin
Defining Citation: PMID:27870838
Proper citation: RRID:Addgene_40775 Copy
Species: Homo sapiens
Genetic Insert: Pin1
Vector Backbone Description: Backbone Size:5200; Vector Backbone:pMCSG7; Vector Types:Bacterial Expression; Bacterial Resistance:Ampicillin
Proper citation: RRID:Addgene_40773 Copy
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