Locus-specific editing of histone modifications at endogenous enhancers.
Mendenhall EM, Williamson KE, Reyon D, Zou JY, Ram O, Joung JK, Bernstein BE
Nat Biotechnol. 2013 Sep 8. doi: 10.1038/nbt.2701.
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PubMed
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Article
The TALE plasmid vectors described on this page are designed to target and edit the epigenome of putative enhancers in mammalian cells. These plasmids allow for cloning DNA fragments encoding TAL effector repeat arrays (assembled using the REAL, REAL-Fast, or FLASH methods) for expression as TALE-LSD1 fusions.
TALE fusions with the histone demethylase LSD1 (Lysine specific demethylase) generated using these plasmids, along with the FLASH method, can decrease the amount of Histone H3 lysine 4 (H3K4) di-methylation at the target locus. This epigenome editing was also shown to alter regulation of nearby gene expression.
The vectors listed below have the EF1α promoter for efficient mammalian cell expression, and encode one of three 0.5 TALE repeats (C, T, and G) and the full length human LSD1 at the carboxy-terminus of the protein.
| ID | Plasmid | Demethylase Domain | C-terminal 0.5 TALE repeat |
|---|---|---|---|
| 49042 |
EMM65
: Backbone contains SHD/C 0.5 Domain TALE
N-terminal: TAL C-terminus: LSD1: Isoform 2 |
LSD1 | HD (binds C) |
| 49043 |
EMM67
: Backbone contains SHD/G 0.5 Domain TALE
N-terminal: TAL C-terminus: LSD1: Isoform 2 |
LSD1 | NN (binds G) |
| 49044 |
EMM71T
: Backbone contains SHD/T 0.5 Domain TALE
N-terminal: TAL C-terminus: LSD1: Isoform 2 |
LSD1 | NG (binds T) |
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 49042 | EMM65 | Backbone contains SHD/C 0.5 Domain TALE N-terminal: TAL C-terminus: LSD1: Isoform 2 |
| 49043 | EMM67 | Backbone contains SHD/G 0.5 Domain TALE N-terminal TALE C-terminus, LSD1: Isoform 2 |
| 49044 | EMM71T | Backbone contains SHD/T 0.5 Domain TALE N-terminal TALE C-terminus, LSD1: Isoform 2 |