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A Novel CRISPR/Cas9 Knock‐in Rat Model Defines A Polymorphic Variant of Secreted Phosphoprotein 2 as A Quantitative Trait Nucleotide Linked to The Heritability of Blood Pressure
Author(s) -
Chakraborty Saroj,
Nie Ying,
Cheng Xi,
Mell Blair,
Galla Sarah,
Joe Bina
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.1026.19
Subject(s) - locus (genetics) , genetics , allele , biology , congenic , candidate gene , spontaneously hypertensive rat , gene , single nucleotide polymorphism , gene knockin , founder effect , genotype , haplotype , blood pressure , endocrinology
Hypertension is a complex polygenic disease caused by a combination of inherited (genetic) and environmental factors. Rat models of inherited hypertension serve as tools to dissect and prioritize genetic factors as candidate genes causing hypertension. One such candidate gene prioritized through systematic linkage and substitution mapping is Spp2 or Secreted Phosphoprotein 2. A single non‐synonymous G/T polymorphism between the Dahl Salt‐ Sensitive (S) rats and Spontaneously Hypertensive Rats (SHR) at the Spp2 locus was hypothesized to cause a reduction in blood pressure (BP) observed in the S.SHR congenic strain spanning the Spp2 locus. To test this hypothesis, a novel rat model was generated using the CRISPR/Cas9 precision‐engineering technology, whereby the ‘G’ allele at the Spp2 locus of the S rat was replaced by the ‘T’ allele of the SHR rat. Protein modeling prediction by SWISSPROT indicated a significantly altered protein structure of the Spp2 protein in the resultant Spp2 knock‐ in rescue model. Following transgenesis, pups born were genotyped and grouped into founders and non‐founders. The founder and non‐founder rats were isogenic except for the Spp2 locus, wherein the founder S rats contained the ‘G’ allele and the non‐founder S rats contained the ‘T’ allele. Both founder and non‐founder rats (n=12 male rats/group) were fed a high salt (2% NaCl) containing diet and their BP was monitored by radiotelemetry. Both systolic and diastolic BP of the Spp2 knock‐in rescue model were significantly lower compared to that of the non‐founder S rats. These data provide conclusive evidence for a single nucleotide polymorphism within the Spp2 gene as a quantitative trait nucleotide (QTN) responsible for the inheritance of blood pressure Support or Funding Information Funding for this work to BJ from the NHLBI/NIH (HL020176) is gratefully acknowledged