@article{Wienholds01122003,
author = {Wienholds, Erno and van Eeden, Freek and Kosters, Marit and Mudde, Josine and Plasterk, Ronald H.A. and Cuppen, Edwin}, 
title = {Efficient Target-Selected Mutagenesis in Zebrafish},
volume = {13}, 
number = {12}, 
pages = {2700-2707}, 
year = {2003}, 
doi = {10.1101/gr.1725103}, 
abstract ={One of the most powerful methods available to assign function to a gene is to inactivate or knockout the gene. Recently, we described the first target-selected knockout in zebrafish. Here, we report on the further improvements of this procedure, resulting in a highly efficient and easy method to do target-selected mutagenesis in zebrafish. A library of 4608 ENU-mutagenized F1 animals was generated and kept as a living stock. The DNA of these animals was screened for mutations in 16 genes by use of CEL-I-mediated heteroduplex cleavage (TILLING) and subsequent resequencing. In total, 255 mutations were identified, of which 14 resulted in a premature stop codon, 7 in a splice donor/acceptor site mutation, and 119 in an amino acid change. By this method, we potentially knocked out 13 different genes in a few months time. Furthermore, we show that TILLING can be used to detect the full spectrum of ENU-induced mutations in a vertebrate genome with the presence of many naturally occurring polymorphisms.}, 
URL = {http://genome.cshlp.org/content/13/12/2700.abstract}, 
eprint = {http://genome.cshlp.org/content/13/12/2700.full.pdf+html}, 
journal = {Genome Research} 
}