When working with TALEN™ in plasmid format, the vector backbone does not replicate and is not prone to integration. Therefore TALEN™ plasmids are typically lost out after several rounds of cell division. However, in order to ensure transient expression of the TALEN™, mRNA delivery method is recommended.
Yes, many publications are already available.
For example, the special edition of the October 2011 nature collections, dedicated to TAL Effector Nucleases, brings together prominent papers recently published on TALENs in Nature biotechnology. A print version can be requested by contacting your local sales manager.
Please click here to access a permanently updated reference list of TALEN publications.
TALEN™ function in a full-range of animal and plant cells and organisms by evoking highly conserved mechanisms of DNA repair. Recent publications on TAL Effector Nucleases provide an overview on their applications and outline gene knock-out and modification in human cell lines and pluripotent cells (hiPS, hES), rat, mous, zebrafish, yeast, C. elegans, C. briggsae, A.thaliana, N. benthamiana.
TALEN™ can be delivered to cells through standard transfection, nucleoporation and micro-injection approaches as either DNA or mRNA. Our plasmid vector backbones allow expression from CMV or EF1a promoters, whilst the T7 promoter allows for in vitro TALEN™ mRNA synthesis and use.
Our vector backbones enable a full-range of delivery protocols, and we recommend working with a method that ensures highest efficiency delivery in your cells of interest.
For technical support, please contact us.
TALEN™ are supplied in a replicable plasmid vector format. Within this vector you have the choice of EF1a or T7 promoters; a dual CMV/T7 promoter; or a promoterless MCS format, suitable for your subcloning strategies.
Yes, TALEN™ have been shown to be able to disrupt multiple copies at once. The efficiency varies on different factors such as cell type, transfection efficiency and TALEN™ activity. We have shown biallelic knock-out in primary human cells at over 50%.
Yes, we have been able to re-target ACTB gene using two different matrices containing two different dyes (GFP and then RFP).
It is important to use different dyes in order to easily identify the retargeting events
That is dependent on cell lines, transfection conditions, and TALEN™ activity. We recommend testing different ratio of Donor Matrix vs. TALEN™ expression vector. For example, in a gene tagging experiment in U-2-OS cells, we used successfully a ratio of 5µg of Donor Matrix and 10µg of each TALEN™ unit.