Profiling the DNA-binding specificities of engineered Cys2His2 zinc finger domains using a rapid cell-based method

@article{citeulike:1421849, abstract = {The C2H2 zinc finger is the most commonly utilized framework for engineering DNA-binding domains with novel specificities. Many different selection strategies have been developed to identify individual fingers that possess a particular DNA-binding specificity from a randomized library. In these experiments, each finger is selected in the context of a constant finger framework that ensures the identification of clones with a desired specificity by properly positioning the randomized finger on the DNA template. Following a successful selection, multiple zinc-finger clones are typically recovered that share similarities in the sequences of their DNA-recognition helices. In principle, each of the clones isolated from a selection is a candidate for assembly into a larger multi-finger protein, but to date a high-throughput method for identifying the most specific candidates for incorporation into a final multi-finger protein has not been available. Here we describe the development of a specificity profiling system that facilitates rapid and inexpensive characterization of engineered zinc-finger modules. Moreover, we demonstrate that specificity data collected using this system can be employed to rationally design zinc fingers with improved DNA-binding specificities. 10.1093/nar/gkm385}, author = {Meng, Xiangdong and Thibodeau-Beganny, Stacey and Jiang, Tao and Joung, Keith J. and Wolfe, Scot A. }, citeulike-article-id = {1421849}, doi = {http://dx.doi.org/10.1093/nar/gkm385}, journal = {Nucl. Acids Res.}, keywords = {zinc\_fingers}, month = {June}, number = {11}, pages = {e81+}, posted-at = {2007-06-29 10:04:28}, priority = {2}, title = {Profiling the DNA-binding specificities of engineered Cys2His2 zinc finger domains using a rapid cell-based method}, url = {http://dx.doi.org/10.1093/nar/gkm385}, volume = {35}, year = {2007} }

TY - JOUR ID - citeulike:1421849 L3 - citeulike-article-id:1421849 N2 - The C2H2 zinc finger is the most commonly utilized framework for engineering DNA-binding domains with novel specificities. Many different selection strategies have been developed to identify individual fingers that possess a particular DNA-binding specificity from a randomized library. In these experiments, each finger is selected in the context of a constant finger framework that ensures the identification of clones with a desired specificity by properly positioning the randomized finger on the DNA template. Following a successful selection, multiple zinc-finger clones are typically recovered that share similarities in the sequences of their DNA-recognition helices. In principle, each of the clones isolated from a selection is a candidate for assembly into a larger multi-finger protein, but to date a high-throughput method for identifying the most specific candidates for incorporation into a final multi-finger protein has not been available. Here we describe the development of a specificity profiling system that facilitates rapid and inexpensive characterization of engineered zinc-finger modules. Moreover, we demonstrate that specificity data collected using this system can be employed to rationally design zinc fingers with improved DNA-binding specificities. 10.1093/nar/gkm385 IS - 11 JF - Nucl. Acids Res. TI - Profiling the DNA-binding specificities of engineered Cys2His2 zinc finger domains using a rapid cell-based method VL - 35 SP - e81 KW - zinc_fingers AU - Meng, Xiangdong AU - Thibodeau-Beganny, Stacey AU - Jiang, Tao AU - Joung, Keith AU - Wolfe, Scot PY - 2007/06/28/ UR - http://dx.doi.org/10.1093/nar/gkm385 ER -