Abstract P042: Development of a Novel Gper-1 Knock Out Rat Model Using a Modified Crispr/cas9 Technology
The G-protein coupled estrogen receptor (Gper-1 or Gpr 30) is a newly recognized estrogen receptor that is widely expressed in various tissues including heart and blood vessels. Rat Gper-1 is a single exonic gene located on chromosome 12. Gper-1 is implicated in the regulation of blood pressure in female mice potentially through its function as a receptor for estrogen. These studies conducted using mice that are without a genetic background permissive for the development of hypertension are not particularly useful for evaluating the function of Gper-1 in the context of hypertension. To understand the function of Gper-1 in the context of a genetically permissive background, we attempted to knock-out the Gper-1 gene on the genome of the Dahl-salt sensitive (S) rat using an modified Clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9 (CRISPR/Cas9) method. To ensure complete knock-out, instead of the traditional method of using a single gRNA, two gRNAs, each targeting one end of the 1128bp Gper-1 gene were developed. The gRNAs were injected into the Dahl S rat embryos. The embryos were then implanted into ten pseudo-pregnant females. Among the 125 pups born, 5 homozygous founders, 21 heterozygotes and 3 partial Gper-1 deletion founders were identified. In conclusion, using an advanced CRISPR/Cas9 technique a panel of Gper-1 rat knock-outs and targeted mutants were generated, which will serve as novel models for studying the structure-function relationships of the Gper-1 gene.
Author Disclosures: H. Waghulde: None. X. Cheng: None. B. Mell: None. S. Miller: None. W. Filipaik: None. T. Saunders: None. B. Joe: None.
- © 2015 by American Heart Association, Inc.