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Addgene inc porange
Porange, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/porange/pORANGE+Cloning+template+vector+(Plasmid+%23131471)/pmc12848079-158-12-13
Average 93 stars, based on 22 article reviews
porange - by Bioz Stars, 2026-09
93/100 stars

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Related Articles

Clone Assay:

Article Title: SHIP164 is a chorein motif lipid transfer protein that controls endosome–Golgi membrane traffic
Article Snippet: .. Briefly, a gRNA targeted close to the desired site within the endogenous reading frame of the indicated target protein was identified and subsequently cloned into empty pORANGE (Addgene plasmid #131471) using HiFi assembly at the tandem BbsI site. ..

Article Title: Regulation of cellular cholesterol distribution via non-vesicular lipid transport at ER-Golgi contact sites.
Article Snippet: .. The CRISPR targeting sites were synthesized by annealing ORP9_ORANGE_4_CT_S and ORP9_ORANGE_4_CT_AS for ORP9, and OSBP_ORANGE_gRNA_2_NT_S and Nature Communications | (2023) 14:5867 18 OSBP_ORANGE_gRNA_2_NT_AS for OSBP, respectively, and individually sub-cloned into pORANGE (Addgene, #131471). cDNA corresponding to the indicated fluorescent protein (mNeonGreen forORP9,mScarletI for OSBP) with a linker region was then cloned into the HindIII and XhoI sites of the pORANGE to generate pORANGE_ORP9_4_CT and pORANGE_mScarlet-I-OSBP_site_2. ..

Article Title: A role for Fis1 in dendritic development
Article Snippet: .. Oligos of each selected guide were ordered through IDT and cloned into pOrange (Addgene plasmid 131471) between Bbs1. ..

Article Title: A role for Fis1 in dendritic development.
Article Snippet: .. Oligos of each selected guide were ordered through IDT and cloned into pOrange (Addgene plasmid 131471) between Bbs1. ..

Article Title: Regulation of cellular cholesterol distribution via non-vesicular lipid transport at ER-Golgi contact sites
Article Snippet: .. The CRISPR targeting sites were synthesized by annealing ORP9_ORANGE_4_CT_S and ORP9_ORANGE_4_CT_AS for ORP9, and OSBP_ORANGE_gRNA_2_NT_S and OSBP_ORANGE_gRNA_2_NT_AS for OSBP, respectively, and individually sub-cloned into pORANGE (Addgene, #131471). cDNA corresponding to the indicated fluorescent protein (mNeonGreen for ORP9, mScarlet-I for OSBP) with a linker region was then cloned into the HindIII and XhoI sites of the pORANGE to generate pORANGE_ORP9_4_CT and pORANGE_mScarlet-I-OSBP_site_2. ..

Article Title: Fis1 is required for the development of the dendritic mitochondrial network in pyramidal cortical neurons
Article Snippet: pCAG GCaMP6f was created by PCR of GCaMP6f from Addgene plasmid 40755 (a gift from Douglas Kim) and cloning it 3’ to the CAG promoter. .. Oligos of each selected guide were ordered through IDT and cloned into pOrange (Addgene plasmid 131471) between Bbs1. .. pCAG erGCaMP6-150 was created by PCR of GCaMP6-150 from Addgene plasmid 91777 (a gift from Douglas Kim) and cloning it 3’ to the CAG promoter while adding a KDEL retention motif to the C-terminus via the primers.

Plasmid Preparation:

Article Title: SHIP164 is a chorein motif lipid transfer protein that controls endosome–Golgi membrane traffic
Article Snippet: .. Briefly, a gRNA targeted close to the desired site within the endogenous reading frame of the indicated target protein was identified and subsequently cloned into empty pORANGE (Addgene plasmid #131471) using HiFi assembly at the tandem BbsI site. ..

Article Title: A role for Fis1 in dendritic development
Article Snippet: .. Oligos of each selected guide were ordered through IDT and cloned into pOrange (Addgene plasmid 131471) between Bbs1. ..

Article Title: Spontaneous and evoked synaptic vesicle release arises from a single releasable pool.
Article Snippet: pLenti-Synapsin-pHoenix-WPRE was a gift from C. Rosenmund (Addgene plasmid #70111). pEGFP-Synaptophysin-pHluorin was a gift from L. Lagnado (Addgene plasmid #24478). .. Lentiviral packaging and envelope vectors pCMVDR8.2 and pMD2.G were a gift from Didier Trono (Addgene plasmid # 12263 and plasmid #12259). pOrange was a gift from Harold MacGillavry (Addgene plasmid #131471) The plasmids coding for Synaptophysin1-mRFP and Synaptobrevin2-pHluorin have been described before.55 pcDNA3Syn-Vti1a-pHluorin was a gift from G. Fischer von Mollard. .. The coding region of pAAV.hSynapsin.SF-iGluSnFR.A184S (gift from Loren Looger, Addgene plasmid # 106174) was amplified by PCR (Primers: iGluSnFR_For 5-GGATCCGCCACCATGGAGACAGA CACACTCCTGC-3/iGluSnFR_ Rev 5-GCGAATTCCTAACGTGGCTTCTTCTGCCAAAG-3) and cloned into amodified pcDNA3 vector containing a Synapsin1 promoter using BamHI and EcoRI restriction sites.

Article Title: A role for Fis1 in dendritic development.
Article Snippet: .. Oligos of each selected guide were ordered through IDT and cloned into pOrange (Addgene plasmid 131471) between Bbs1. ..

Article Title: Fis1 is required for the development of the dendritic mitochondrial network in pyramidal cortical neurons
Article Snippet: pCAG GCaMP6f was created by PCR of GCaMP6f from Addgene plasmid 40755 (a gift from Douglas Kim) and cloning it 3’ to the CAG promoter. .. Oligos of each selected guide were ordered through IDT and cloned into pOrange (Addgene plasmid 131471) between Bbs1. .. pCAG erGCaMP6-150 was created by PCR of GCaMP6-150 from Addgene plasmid 91777 (a gift from Douglas Kim) and cloning it 3’ to the CAG promoter while adding a KDEL retention motif to the C-terminus via the primers.

CRISPR:

Article Title: Regulation of cellular cholesterol distribution via non-vesicular lipid transport at ER-Golgi contact sites.
Article Snippet: .. The CRISPR targeting sites were synthesized by annealing ORP9_ORANGE_4_CT_S and ORP9_ORANGE_4_CT_AS for ORP9, and OSBP_ORANGE_gRNA_2_NT_S and Nature Communications | (2023) 14:5867 18 OSBP_ORANGE_gRNA_2_NT_AS for OSBP, respectively, and individually sub-cloned into pORANGE (Addgene, #131471). cDNA corresponding to the indicated fluorescent protein (mNeonGreen forORP9,mScarletI for OSBP) with a linker region was then cloned into the HindIII and XhoI sites of the pORANGE to generate pORANGE_ORP9_4_CT and pORANGE_mScarlet-I-OSBP_site_2. ..

Article Title: Regulation of cellular cholesterol distribution via non-vesicular lipid transport at ER-Golgi contact sites
Article Snippet: .. The CRISPR targeting sites were synthesized by annealing ORP9_ORANGE_4_CT_S and ORP9_ORANGE_4_CT_AS for ORP9, and OSBP_ORANGE_gRNA_2_NT_S and OSBP_ORANGE_gRNA_2_NT_AS for OSBP, respectively, and individually sub-cloned into pORANGE (Addgene, #131471). cDNA corresponding to the indicated fluorescent protein (mNeonGreen for ORP9, mScarlet-I for OSBP) with a linker region was then cloned into the HindIII and XhoI sites of the pORANGE to generate pORANGE_ORP9_4_CT and pORANGE_mScarlet-I-OSBP_site_2. ..

Synthesized:

Article Title: Regulation of cellular cholesterol distribution via non-vesicular lipid transport at ER-Golgi contact sites.
Article Snippet: .. The CRISPR targeting sites were synthesized by annealing ORP9_ORANGE_4_CT_S and ORP9_ORANGE_4_CT_AS for ORP9, and OSBP_ORANGE_gRNA_2_NT_S and Nature Communications | (2023) 14:5867 18 OSBP_ORANGE_gRNA_2_NT_AS for OSBP, respectively, and individually sub-cloned into pORANGE (Addgene, #131471). cDNA corresponding to the indicated fluorescent protein (mNeonGreen forORP9,mScarletI for OSBP) with a linker region was then cloned into the HindIII and XhoI sites of the pORANGE to generate pORANGE_ORP9_4_CT and pORANGE_mScarlet-I-OSBP_site_2. ..

Article Title: Regulation of cellular cholesterol distribution via non-vesicular lipid transport at ER-Golgi contact sites
Article Snippet: .. The CRISPR targeting sites were synthesized by annealing ORP9_ORANGE_4_CT_S and ORP9_ORANGE_4_CT_AS for ORP9, and OSBP_ORANGE_gRNA_2_NT_S and OSBP_ORANGE_gRNA_2_NT_AS for OSBP, respectively, and individually sub-cloned into pORANGE (Addgene, #131471). cDNA corresponding to the indicated fluorescent protein (mNeonGreen for ORP9, mScarlet-I for OSBP) with a linker region was then cloned into the HindIII and XhoI sites of the pORANGE to generate pORANGE_ORP9_4_CT and pORANGE_mScarlet-I-OSBP_site_2. ..



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F-actin influences the trafficking of GS in axons. a ) Representative confocal images of Golgi satellites in proximal axons following latrunculin A (LatA) treatment (5 µM, right) and DMSO control treatment (left) in relation to <t>actin</t> <t>(pORANGE-actin-tagRFP</t> endogenous labelling and <t>St3gal5-GFP</t> overexpression). See also videos 4 (DMSO) and 5 (LatA). b ) Kymographs of the videos shown in a), recorded for 3 min at 2 FPS, showing increased movement and decreased pausing of the GS after LatA treatment compared to DMSO ctr. c ) Quantification of GS trafficking parameters in the proximal axon: run length, velocity, pausing time and stationary GS; run length and velocity are increased in the LatA treatment group (p (anterograde run length) = 0.0891, p* (retrograde run length) = 0.0455, p* (anterograde velocity) = 0.0425, p* (retrograde velocity = 0.0334); unpaired t-test; n (DMSO) = 9, n (LatA) = 10; one axon from each cell in 3 individual cultures). The pausing time is significantly reduced by LatA treatment (p* = 0.0240; unpaired t-test; n (DMSO) = 15, n (LatA) = 14; one axon from each cell in 3 individual cultures). All values are presented as mean ± SD. Quantification of stationary events per 10 μm showing no difference in stationary events ( p = 0.9659; unpaired t-test; n (DMSO) = 15, n (LatA) = 14; one axon from one cell in 3 single cultures), presented as mean ± SD. d ) Representative kymographs of confocal time-lapse images of GS in proximal axons following 30 min of latrunculin A (LatA, 5 µM, right panel) and DMSO control treatment (left). Videos and Kymographs as described in a) and b). e ) Quantification of immobile GS in the distal axon: pausing time and stationary events are measured. The pausing time is significantly reduced by LatA treatment (p* = 0,0490; unpaired t-test; n (DMSO) = 14, n (LatA) = 16; one axon from each cell in 3 individual cultures). All values are presented as mean ± SD. Quantification of stationary events per 10 μm being significantly reduced by LatA treatment (p* = 0.0274; unpaired t-test n (DMSO) = 14, n (LatA) = 16; one axon from one cell in 3 single cultures), presented as mean ± SD
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Image Search Results


F-actin influences the trafficking of GS in axons. a ) Representative confocal images of Golgi satellites in proximal axons following latrunculin A (LatA) treatment (5 µM, right) and DMSO control treatment (left) in relation to actin (pORANGE-actin-tagRFP endogenous labelling and St3gal5-GFP overexpression). See also videos 4 (DMSO) and 5 (LatA). b ) Kymographs of the videos shown in a), recorded for 3 min at 2 FPS, showing increased movement and decreased pausing of the GS after LatA treatment compared to DMSO ctr. c ) Quantification of GS trafficking parameters in the proximal axon: run length, velocity, pausing time and stationary GS; run length and velocity are increased in the LatA treatment group (p (anterograde run length) = 0.0891, p* (retrograde run length) = 0.0455, p* (anterograde velocity) = 0.0425, p* (retrograde velocity = 0.0334); unpaired t-test; n (DMSO) = 9, n (LatA) = 10; one axon from each cell in 3 individual cultures). The pausing time is significantly reduced by LatA treatment (p* = 0.0240; unpaired t-test; n (DMSO) = 15, n (LatA) = 14; one axon from each cell in 3 individual cultures). All values are presented as mean ± SD. Quantification of stationary events per 10 μm showing no difference in stationary events ( p = 0.9659; unpaired t-test; n (DMSO) = 15, n (LatA) = 14; one axon from one cell in 3 single cultures), presented as mean ± SD. d ) Representative kymographs of confocal time-lapse images of GS in proximal axons following 30 min of latrunculin A (LatA, 5 µM, right panel) and DMSO control treatment (left). Videos and Kymographs as described in a) and b). e ) Quantification of immobile GS in the distal axon: pausing time and stationary events are measured. The pausing time is significantly reduced by LatA treatment (p* = 0,0490; unpaired t-test; n (DMSO) = 14, n (LatA) = 16; one axon from each cell in 3 individual cultures). All values are presented as mean ± SD. Quantification of stationary events per 10 μm being significantly reduced by LatA treatment (p* = 0.0274; unpaired t-test n (DMSO) = 14, n (LatA) = 16; one axon from one cell in 3 single cultures), presented as mean ± SD

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Myosin VI controls localization of golgi satellites at active presynaptic boutons

doi: 10.1007/s00018-025-05896-2

Figure Lengend Snippet: F-actin influences the trafficking of GS in axons. a ) Representative confocal images of Golgi satellites in proximal axons following latrunculin A (LatA) treatment (5 µM, right) and DMSO control treatment (left) in relation to actin (pORANGE-actin-tagRFP endogenous labelling and St3gal5-GFP overexpression). See also videos 4 (DMSO) and 5 (LatA). b ) Kymographs of the videos shown in a), recorded for 3 min at 2 FPS, showing increased movement and decreased pausing of the GS after LatA treatment compared to DMSO ctr. c ) Quantification of GS trafficking parameters in the proximal axon: run length, velocity, pausing time and stationary GS; run length and velocity are increased in the LatA treatment group (p (anterograde run length) = 0.0891, p* (retrograde run length) = 0.0455, p* (anterograde velocity) = 0.0425, p* (retrograde velocity = 0.0334); unpaired t-test; n (DMSO) = 9, n (LatA) = 10; one axon from each cell in 3 individual cultures). The pausing time is significantly reduced by LatA treatment (p* = 0.0240; unpaired t-test; n (DMSO) = 15, n (LatA) = 14; one axon from each cell in 3 individual cultures). All values are presented as mean ± SD. Quantification of stationary events per 10 μm showing no difference in stationary events ( p = 0.9659; unpaired t-test; n (DMSO) = 15, n (LatA) = 14; one axon from one cell in 3 single cultures), presented as mean ± SD. d ) Representative kymographs of confocal time-lapse images of GS in proximal axons following 30 min of latrunculin A (LatA, 5 µM, right panel) and DMSO control treatment (left). Videos and Kymographs as described in a) and b). e ) Quantification of immobile GS in the distal axon: pausing time and stationary events are measured. The pausing time is significantly reduced by LatA treatment (p* = 0,0490; unpaired t-test; n (DMSO) = 14, n (LatA) = 16; one axon from each cell in 3 individual cultures). All values are presented as mean ± SD. Quantification of stationary events per 10 μm being significantly reduced by LatA treatment (p* = 0.0274; unpaired t-test n (DMSO) = 14, n (LatA) = 16; one axon from one cell in 3 single cultures), presented as mean ± SD

Article Snippet: The pORANGE_Actin was produced by exchanging the GFP in the Addgene #139,666 vector (pORANGE_Actin#2-GFP) for a TagRFP (amplified from pTagRFP-C, Evrogen) using the restriction enzymes NheI and BamHI.

Techniques: Control, Over Expression