title: Targeting of the Plzf Gene in the Rat by Transcription Activator-Like Effector Nuclease Results in Caudal Regression Syndrome in Spontaneously Hypertensive Rats.
abstract: Recently, it has been found that spontaneous mutation Lx (polydactyly-luxate syndrome) in the rat is determined by deletion of a conserved intronic sequence of the Plzf (Promyelocytic leukemia zinc finger protein) gene. In addition, Plzf is a prominent candidate gene for quantitative trait loci (QTLs) associated with cardiac hypertrophy and fibrosis in the spontaneously hypertensive rat (SHR). In the current study, we tested the effects of Plzf gene targeting in the SHR using TALENs (transcription activator-like effector nucleases). SHR ova were microinjected with constructs pTAL438/439 coding for a sequence-specific endonuclease that binds to target sequence in the first coding exon of the Plzf gene. Out of 43 animals born after microinjection, we detected a single male founder. Sequence analysis revealed a deletion of G that resulted in frame shift mutation starting in codon 31 and causing a premature stop codon at position of amino acid 58. The Plzftm1Ipcv allele is semi-lethal since approximately 95% of newborn homozygous animals died perinatally. All homozygous animals exhibited manifestations of a caudal regression syndrome including tail anomalies and serious size reduction and deformities of long bones, and oligo- or polydactyly on the hindlimbs. The heterozygous animals only exhibited the tail anomalies. Impaired development of the urinary tract was also revealed: one homozygous and one heterozygous rat exhibited a vesico-ureteric reflux with enormous dilatation of ureters and renal pelvis. In the homozygote, this was combined with a hypoplastic kidney. These results provide evidence for the important role of Plzf gene during development of the caudal part of a body-column vertebrae, hindlimbs and urinary system in the rat.
authors.affiliation: Institute of Biology and Medical Genetics, 1st Medical Faculty, Charles University in Prague, Prague, Czech Republic.
authors.forename: František
authors.initials: F
authors.lastname: Liška
authors.affiliation: Institute of Experimental Medicine, Czech Academy of Sciences, Prague, Czech Republic.
authors.forename: Renata
authors.initials: R
authors.lastname: Peterková
authors.affiliation: Institute of Experimental Medicine, Czech Academy of Sciences, Prague, Czech Republic.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Peterka
authors.affiliation: Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic.
authors.forename: Vladimír
authors.initials: V
authors.lastname: Landa
authors.affiliation: Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic.
authors.forename: Václav
authors.initials: V
authors.lastname: Zídek
authors.affiliation: Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Mlejnek
authors.affiliation: Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Šilhavý
authors.affiliation: Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic.
authors.forename: Miroslava
authors.initials: M
authors.lastname: Šimáková
authors.affiliation: Institute of Biology and Medical Genetics, 1st Medical Faculty, Charles University in Prague, Prague, Czech Republic.|Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic.
authors.forename: Vladimír
authors.initials: V
authors.lastname: Křen
authors.affiliation: University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Colby G
authors.initials: CG
authors.lastname: Starker
authors.affiliation: University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Voytas
authors.affiliation: Max Delbrück Center for Molecular Medicine, Berlin, Germany.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Izsvák
authors.affiliation: Institute of Biology and Medical Genetics, 1st Medical Faculty, Charles University in Prague, Prague, Czech Republic.|Institute of Experimental Medicine, Czech Academy of Sciences, Prague, Czech Republic.
authors.forename: Michal
authors.initials: M
authors.lastname: Pravenec
mesh_terms: D000015:Abnormalities, Multiple / Q000235:genetics / Q000473:pathology / Q000662:veterinary; D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D005091:Exons; D016368:Frameshift Mutation; D018390:Gene Targeting; D005838:Genotype; D006579:Heterozygote; D006720:Homozygote; D008297:Male; D017689:Polydactyly / Q000235:genetics / Q000473:pathology / Q000662:veterinary; D000075722:Promyelocytic Leukemia Zinc Finger Protein; D011485:Protein Binding; D040641:Quantitative Trait Loci; D051381:Rats; D011918:Rats, Inbred SHR; D013623:Tail / Q000002:abnormalities; D000069896:Transcription Activator-Like Effector Nucleases / Q000235:genetics / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D000075722:Promyelocytic Leukemia Zinc Finger Protein; C534029:ZBTB16 protein, rat; D000069896:Transcription Activator-Like Effector Nucleases