A Transcription Activator-Like Effector (TALE) Toolbox for Genome Engineering

By Neville E. Sanjana, Le Cong, Yang Zhou, Margaret M Cunniff, G. Feng, Feng Zhang. Nature Protocols, 2012

670 citations recorded by the source corpus.

Transcription activator-like effectors (TALEs) are a class of naturally occurring DNA-binding proteins found in the plant pathogen Xanthomonas sp. The DNA-binding domain of each TALE consists of tandem 34–amino acid repeat modules that can be rearranged according to a simple cipher to target new DNA sequences. Customized TALEs can be used for a wide variety of genome engineering applications, including transcriptional modulation and genome editing. Here we describe a toolbox for rapid construction of custom TALE transcription factors (TALE-TFs) and nucleases (TALENs) using a hierarchical ligation procedure. This toolbox facilitates affordable and rapid construction of custom TALE-TFs and TALENs within 1 week and can be easily scaled up to construct TALEs for multiple targets in parallel. We also provide details for testing the activity in mammalian cells of custom TALE-TFs and TALENs using quantitative reverse-transcription PCR and Surveyor nuclease, respectively. The TALE toolbox described here will enable a broad range of biological applications.

Subfields: Molecular Biology, Plant Science

View Paper: https://www.semanticscholar.org/paper/d6b9fd53e8760512d49c202c0cb7545c96db0dbb

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Molecular Biology

In Plant Science

Citations in the corpus

CRISPR interference (CRISPRi) for sequence-specific control of gene expression (2013, background): “Several targeted gene regulation techniques have been widely used in the past, such as RNA interference (RNAi) 27,28 or engineered DNA-binding proteins, including zinc-finger 29–31 or transcription activator–like effector (TALE) 32–34 proteins.”
A single oncogenic enhancer rearrangement causes concomitant EVI1 and GATA2 deregulation in leukemia. (2014, background): “…18 kbCTSPhysically Interact A hallmark of distal enhancer elements is their engagement in chromatin loops physically contacting with promoters to induce transcription factor assembly and polymerase II recruitment (Deng et al., 2012; Sanyal et al., 2012; Thurman et al., 2012; Tolhuis et al., 2002).”
RNA-Guided Human Genome Engineering via Cas9 (2013, positive): “TALENs used in this study were constructed using the protocols described in (11).”
Optimization of scarless human stem cell genome editing (2013, methodology): “re-TALENs and re-TALE-TF destination vectors were constructed by modifying the TALE-TF and TALEN cloning backbones (24).”
Genome Engineering of Crops with Designer Nucleases (2012, methodology): “…dTALEs. h ey include Golden Gate assembly methods that allow seamless construction of TALE repeat arrays (Cermak et al., 2011; Engler et al., 2009; Sanjana et al., 2012; Zhang et al., 2011) as well as ligation-based systems (Reyon et al., 2012; Sander et al., 2011a). h ese platforms make it…”
Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity (2013, background): “…effector nucleases (TALENs) (Boch et al., 2009; Christian et al., 2010; Miller et al., 2011; Moscou and Bogdanove, 2009; Reyon et al., 2012; Sanjana et al., 2012; Wood et al., 2011; Zhang et al., 2011), wherein DNA cleavage requires synergistic interaction of two independent…”