RT Journal
A1 Spielmann, Malte
A1 Kakar, Naseebullah
A1 Tayebi, Naeimeh
A1 Leettola, Catherine
A1 Nürnberg, Gudrun
A1 Sowada, Nadine
A1 Lupiáñez, Darío G.
A1 Harabula, Izabela
A1 Flöttmann, Ricarda
A1 Horn, Denise
A1 Chan, Wing Lee
A1 Wittler, Lars
A1 Yilmaz, Rüstem
A1 Altmüller, Janine
A1 Thiele, Holger
A1 van Bokhoven, Hans
A1 Schwartz, Charles E.
A1 Nürnberg, Peter
A1 Bowie, James U.
A1 Ahmad, Jamil
A1 Kubisch, Christian
A1 Mundlos, Stefan
A1 Borck, Guntram
T1 Exome sequencing and CRISPR/Cas genome editing identify mutations of ZAK as a cause of limb defects in humans and mice
JF Genome Research 
JO Genome Research 
YR 2016 
FD February 01 
VO 26 
IS 2 
SP 183 
OP 191 
DO 10.1101/gr.199430.115 
UL http://genome.cshlp.org/content/26/2/183.abstract 
AB The CRISPR/Cas technology enables targeted genome editing and the rapid generation of transgenic animal models for the study of human genetic disorders. Here we describe an autosomal recessive human disease in two unrelated families characterized by a split-foot defect, nail abnormalities of the hands, and hearing loss, due to mutations disrupting the SAM domain of the protein kinase ZAK. ZAK is a member of the MAPKKK family with no known role in limb development. We show that Zak is expressed in the developing limbs and that a CRISPR/Cas-mediated knockout of the two Zak isoforms is embryonically lethal in mice. In contrast, a deletion of the SAM domain induces a complex hindlimb defect associated with down-regulation of Trp63, a known split-hand/split-foot malformation disease gene. Our results identify ZAK as a key player in mammalian limb patterning and demonstrate the rapid utility of CRISPR/Cas genome editing to assign causality to human mutations in the mouse in <10 wk.