pdCas9-DNMT3A-EGFP (ANV) | Addgene 71685 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 :

pdCas9-DNMT3A-EGFP (ANV)

catalog :

71685

citations: 8

Reference
Kehayova Y, Wilkinson J, Rice S, Loughlin J. Independent osteoarthritis risk-conferring alleles mediate the same epigenetic and transcriptional effect on a shared target gene, COLGALT2. Arthritis Rheumatol. 2022;: pubmed publisher
Singh K, Rustagi Y, Abouhashem A, Tabasum S, Verma P, Hernandez E, et al. Genome-wide DNA hypermethylation opposes healing in patients with chronic wounds by impairing epithelial-mesenchymal transition. J Clin Invest. 2022;132: pubmed publisher
Truong V, Lin Y, Nguyen N, Hsu M, Pham N, Chang Y, et al. Bi-directional gene activation and repression promote ASC differentiation and enhance bone healing in osteoporotic rats. Mol Ther. 2022;30:92-104 pubmed publisher
Li M, Jiang P, Cheng K, Zhang Z, Lan S, Li X, et al. Regulation of MYB by distal enhancer elements in human myeloid leukemia. Cell Death Dis. 2021;12:223 pubmed publisher
Qian P, De Kumar B, He X, Nolte C, Gogol M, Ahn Y, et al. Retinoid-Sensitive Epigenetic Regulation of the Hoxb Cluster Maintains Normal Hematopoiesis and Inhibits Leukemogenesis. Cell Stem Cell. 2018;22:740-754.e7 pubmed publisher
Galonska C, Charlton J, Mattei A, Donaghey J, Clement K, Gu H, et al. Genome-wide tracking of dCas9-methyltransferase footprints. Nat Commun. 2018;9:597 pubmed publisher
Xiong T, Meister G, Workman R, Kato N, Spellberg M, Turker F, et al. Targeted DNA methylation in human cells using engineered dCas9-methyltransferases. Sci Rep. 2017;7:6732 pubmed publisher
Vojta A, DobriniÄ‡ P, TadiÄ‡ V, BoÄkor L, KoraÄ‡ P, Julg B, et al. Repurposing the CRISPR-Cas9 system for targeted DNA methylation. Nucleic Acids Res. 2016;44:5615-28 pubmed publisher

product information

Catalog Number :

71685

Product Name :

pdCas9-DNMT3A-EGFP (ANV)

article :

doi	10.1093/nar/gkw159
id	18013
pubmed_id	26969735

bacterial resistance :

Ampicillin

cloning :

backbone

PX462

backbone_mutation

backbone_origin

backbone_size

4246

promoter

sequencing_primer_3

sequencing_primer_5

vector_types

Mammalian Expression

CRISPR

growth notes :

The catalytic domain of human DNMT3A (amino acids P602-V912) was derived from the plasmid pcDNA3/Myc-DNMT3A (Addgene, plasmid #35521) (Chen et al., 2005, J Cell Biochem 95: 902-917). Undesired BbsI restriction site was removed by site-directed mutagenesis, without affecting the amino acid sequence. The DNMT3A active site motif ENV was mutated to ANV (E756A). Plasmid pSpCas9n(BB)-2A-Puro (PX462) (Addgene, plasmid #48141) (Ran et al., 2013, Nat Protoc 8: 2281-2308) was used as a backbone. Additional H840A mutation was introduced into Cas9n D10A nickase. T2A-PuroR EcoRI fragment was replaced with T2A-EGFP fragment from pSpCas9n(BB)-2A-GFP (PX461) (Addgene, plasmid #48140) (Ran et al., 2013, Nat Protoc 8: 2281-2308).

growth strain :

Expression of mutant human codon-optimized dCas9-DNMT3A fusion (inactive catalytic domain of DNMT3A) with T2A-EGFP; cloning backbone for sgRNA

origin :

pi :

alt_names

dCas9-DNMT3A-EGFP (ANV)

SpdCas9-DNMT3A-T2A-EGFP (ANV)

6020

cloning

clone_method	Restriction Enzyme
cloning_site_3	FseI
cloning_site_5	BamHI
promoter	CBh
sequencing_primer_3	BGH Rev
sequencing_primer_5	GS-linker For 5' GAAGAGGTACACCAGCACCAAAG 3'
site_3_destroyed
site_5_destroyed

entrez_gene

aliases	DNMT3A2, HESJAS, M.HsaIIIA, TBRS
gene	DNMT3A
id	1788

genbank_ids

AKA60242.1

NP_072046.2

mutation

D10A and H840A in S.pyogenes Cas9; E756A inactivating mutation in H. sapiens DNMT3A

name

S. pyogenes dCas9 fused with the inactivated (E756A) catalytic domain of human DNMT3A (amino acids P602-V912) and T2A-EGFP

shRNA_sequence

size

species

9606

Homo sapiens

100

Synthetic

32644

Other

S. pyogenes