Hyperhomocysteinemia and Dyslipidemia in point mutation G307S of cystathionine β-synthase-deficient rabbit generated using CRISPR/Cas9
Background: Congenital hyper-homocysteinemia (HHcy) is caused by a defective cystathionine β-synthase (CBS) gene, and is frequently associated with dyslipdemia. The aim of this study was to further elucidate the effect of mutated CBS gene on circulating lipids using a rabbit model harboring a homozygous G307S point mutation in CBS.
Methods: CRISPR/Cas9 system was used to edit the CBS gene in rabbit embryos. The founder rabbits were sequenced, and their plasma homocysteine (Hcy) and lipid profile were analyzed.
Results: Six CBS-knockout (CBS-KO) founder lines with biallelic modifications were obtained. Mutation in CBS caused significant growth retardation and high mortality rates within 6 weeks after birth. In addition, the 6-week old CBS-KO rabbits showed higher plasma levels of Hcy, triglycerides (TG), total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) compared to the age-matched wild-type (WT) controls. Histological analysis of the mutants showed accumulation of micro-vesicular cytoplasmic lipid droplets in the hepatocytes. However, gastric infusion of vitamin B and betaine complex significantly decreased the plasma levels of TG, TC and LDL-C in the CBS-KO rabbits, and alleviated hepatic steatosis compared to the untreated animals.
Conclusion: A CBSG307S rabbit model was generated that exhibited severe dyslipidemia when fed on a normal diet, indicating that G307S mutation in the CBS gene is a causative factor for dyslipidemia.
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Table S1. The sequences of potential off-target sites. Table S2. The sequences of potential off-target loci PCR primers.
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Hyperhomocysteinemia and Dyslipidemia in point mutation G307S of cystathionine β-synthase-deficient rabbit generated using CRISPR/Cas9
Posted 22 Sep, 2020
On 14 Oct, 2020
Received 23 Sep, 2020
On 23 Sep, 2020
Invitations sent on 22 Sep, 2020
On 22 Sep, 2020
On 21 Sep, 2020
On 20 Sep, 2020
On 20 Sep, 2020
Received 19 Sep, 2020
On 19 Sep, 2020
On 18 Sep, 2020
Received 17 Sep, 2020
Received 15 Sep, 2020
Received 14 Sep, 2020
On 14 Sep, 2020
Invitations sent on 14 Sep, 2020
On 14 Sep, 2020
On 14 Sep, 2020
On 14 Sep, 2020
On 14 Sep, 2020
Received 14 Sep, 2020
On 13 Sep, 2020
On 13 Sep, 2020
On 18 Aug, 2020
Received 13 Aug, 2020
On 11 Aug, 2020
Received 08 Aug, 2020
Received 08 Aug, 2020
Received 08 Aug, 2020
Received 08 Aug, 2020
Invitations sent on 02 Aug, 2020
On 02 Aug, 2020
On 02 Aug, 2020
On 02 Aug, 2020
On 02 Aug, 2020
On 31 Jul, 2020
On 30 Jul, 2020
On 30 Jul, 2020
On 30 Jul, 2020
Background: Congenital hyper-homocysteinemia (HHcy) is caused by a defective cystathionine β-synthase (CBS) gene, and is frequently associated with dyslipdemia. The aim of this study was to further elucidate the effect of mutated CBS gene on circulating lipids using a rabbit model harboring a homozygous G307S point mutation in CBS.
Methods: CRISPR/Cas9 system was used to edit the CBS gene in rabbit embryos. The founder rabbits were sequenced, and their plasma homocysteine (Hcy) and lipid profile were analyzed.
Results: Six CBS-knockout (CBS-KO) founder lines with biallelic modifications were obtained. Mutation in CBS caused significant growth retardation and high mortality rates within 6 weeks after birth. In addition, the 6-week old CBS-KO rabbits showed higher plasma levels of Hcy, triglycerides (TG), total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) compared to the age-matched wild-type (WT) controls. Histological analysis of the mutants showed accumulation of micro-vesicular cytoplasmic lipid droplets in the hepatocytes. However, gastric infusion of vitamin B and betaine complex significantly decreased the plasma levels of TG, TC and LDL-C in the CBS-KO rabbits, and alleviated hepatic steatosis compared to the untreated animals.
Conclusion: A CBSG307S rabbit model was generated that exhibited severe dyslipidemia when fed on a normal diet, indicating that G307S mutation in the CBS gene is a causative factor for dyslipidemia.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5