Hu, Ruozhen Wallace, Jared J. Dahlem, Timothy Jonah Grunwald, David M. O'Connell, Ryan Figure S1 - Targeting Human MicroRNA Genes Using Engineered Tal-Effector Nucleases (TALENs) <p><b>Experimental plan used to develop miRNA-targeting TALENs.</b> TALEN pairs targeting different miRNAs were designed, constructed and transfected into 293T cells along with a GFP expression vector. After transfection, 293T cells were subjected to FACS sort to isolate cells with the TALEN pairs. gDNA was extracted from Wt, unsorted, GFP- or GFP+ cells. The TALEN pair-targeted regions were amplified by PCR and subjected to HRMA or TOPO cloning and sequencing to determine the presence of mutations. Moreover, GFP+ cells were plated in 96 well plates to obtain single cell clones. Single cell clones were subjected to PCR and TOPO cloning analyses to determine if bi- or mono-allelic mutations were being generated within the TALEN targeted regions.</p> <p>(TIF)</p> Biochemistry;Nucleic acids;rna;RNA interference;biotechnology;Genetic engineering;Computational biology;genomics;Functional genomics;Molecular genetics;Gene regulation;gene expression;genetics;Genetic mutation;mutagenesis;Gene function;Molecular cell biology;microrna;genes;engineered;tal-effector;nucleases 2013-05-07
    https://plos.figshare.com/articles/figure/_Targeting_Human_MicroRNA_Genes_Using_Engineered_Tal_Effector_Nucleases_TALENs_/699114
10.1371/journal.pone.0063074.s001