AICSDP-15:ACTB-mEGFP | Addgene 87425 product information

This webpage contains legacy information. The product is either no longer available from the supplier or has been delisted at Labome.

product summary

company name :

Addgene

product type :

cDNA

product name :

AICSDP-15:ACTB-mEGFP

catalog :

87425

citations: 7

Reference
Chang J, Wang C, Lin S. Interrogation of human microglial phagocytosis by CRISPR genome editing. Front Immunol. 2023;14:1169725 pubmed publisher
Bodai Z, Bishop A, Gantz V, Komor A. Targeting double-strand break indel byproducts with secondary guide RNAs improves Cas9 HDR-mediated genome editing efficiencies. Nat Commun. 2022;13:2351 pubmed publisher
Sun D, Evans L, Perrone F, Sokleva V, Lim K, Rezakhani S, et al. A functional genetic toolbox for human tissue-derived organoids. elife. 2021;10: pubmed publisher
Huang R, Lai M, Shih H, Lin S. A robust platform for expansion and genome editing of primary human natural killer cells. J Exp Med. 2021;218: pubmed publisher
Cruz Becerra G, Kadonaga J. Enhancement of homology-directed repair with chromatin donor templates in cells. elife. 2020;9: pubmed publisher
Garner R, Skariah G, Hadjitheodorou A, Belliveau N, Savinov A, Footer M, et al. Neutrophil-like HL-60 cells expressing only GFP-tagged β-actin exhibit nearly normal motility. Cytoskeleton (Hoboken). 2020;77:181-196 pubmed publisher
Roberts B, Haupt A, Tucker A, Grancharova T, Arakaki J, Fuqua M, et al. Systematic gene tagging using CRISPR/Cas9 in human stem cells to illuminate cell organization. Mol Biol Cell. 2017;28:2854-2874 pubmed publisher

product information

Catalog Number :

87425

Product Name :

AICSDP-15:ACTB-mEGFP

article :

doi	10.1091/mbc.E17-03-0209
id	28192202
pubmed_id	28814507

bacterial resistance :

Ampicillin

cloning :

backbone

pUC57

backbone_mutation

backbone_origin

backbone_size

promoter

sequencing_primer_3

sequencing_primer_5

vector_types

Mammalian Expression

CRISPR

Other

Donor Template

growth notes :

This plasmid has been used with locus-specific CRISPR/Cas9 to add a mEGFP tag to the N-terminus of human ACTB in WTC human induced pluripotent stem cells by the Allen Institute for Cell Science. Linker (AA) sequence: AGSGT. After protein tagging using this donor template plasmid and CRISPR/Cas9 reagents, transfected cells may exhibit varying intensity levels of fluorescence, likely due to editing precision. To obtain cells of uniform intensity levels, see our protocol for fluorescence-assisted cell sorting and subcloning of transfected cells (https://www.allencell.org/instructional-videos-and-tutorials-for-cell-methods.html). Further, we recommend PCR-based assays for identifying precisely edited clones as previously described (https://www.molbiolcell.org/doi/abs/10.1091/mbc.e17-03-0209). For more information on the entire plasmid collection, please see https://www.addgene.org/allen-institute-cell-science/ . Please visit https://www.biorxiv.org/content/early/2017/03/31/123042 for bioRxiv preprint.

growth strain :

Homology arms and linker-mEGFP sequence for N-terminus tagging of human ACTB

origin :

pi :

alt_names

ACTB

cloning

clone_method	Unknown
cloning_site_3
cloning_site_5
promoter
sequencing_primer_3	M13-Rev
sequencing_primer_5	M13-F20
site_3_destroyed
site_5_destroyed

entrez_gene

aliases	BKRNS, BNS, BRWS1, CSMH, DDS1, PS1TP5BP1, THC8
gene	ACTB
id	60

genbank_ids

mutation

homology arms contain point mutations to disrupt crRNA binding sites used and point mutations specific to WTC genome

name

ACTB Homology Arms with mEGFP-linker

shRNA_sequence

size

2729

species

9606

Homo sapiens