pdCas9-DNMT3A-PuroR (ANV) | Addgene 71684 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-PuroR (ANV)

catalog :

71684

citations: 5

Reference
Tiane A, Schepers M, Reijnders R, van Veggel L, Chenine S, Rombaut B, et al. From methylation to myelination: epigenomic and transcriptomic profiling of chronic inactive demyelinated multiple sclerosis lesions. Acta Neuropathol. 2023;146:283-299 pubmed publisher
Metzger D, Miller K, Lyon W, Migliozzi R, Pangburn H, Saldanha R. Host Cell Transcriptional Tuning with CRISPR/dCas9 to Mitigate the Effects of Toxin Exposure. ACS Synth Biol. 2022;11:3657-3668 pubmed publisher
Tiane A, Schepers M, Riemens R, Rombaut B, Vandormael P, Somers V, et al. DNA methylation regulates the expression of the negative transcriptional regulators ID2 and ID4 during OPC differentiation. Cell Mol Life Sci. 2021;78:6631-6644 pubmed publisher
Cali C, Park D, Lee E. Targeted DNA methylation of neurodegenerative disease genes via homology directed repair. Nucleic Acids Res. 2019;47:11609-11622 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 :

71684

Product Name :

pdCas9-DNMT3A-PuroR (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.

growth strain :

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

origin :

pi :

alt_names

dCas9-DNMT3A-PuroR (ANV)

SpdCas9-DNMT3A-T2A-PuroR (ANV)

dCas9-DNMT3A (ANV)

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-PuroR

shRNA_sequence

size

5906

species

9606

Homo sapiens

100

Synthetic

32644

Other

S. pyogenes