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Generation of esr1-knockout rats using zinc finger nuclease-mediated genome editing.

Authors: Rumi, MA  Dhakal, P  Kubota, K  Chakraborty, D  Lei, T  Larson, MA  Wolfe, MW  Roby, KF  Vivian, JL  Soares, MJ 
Citation: Rumi MA, etal., Endocrinology. 2014 May;155(5):1991-9. doi: 10.1210/en.2013-2150. Epub 2014 Feb 7.
Pubmed: (View Article at PubMed) PMID:24506075
DOI: Full-text: DOI:10.1210/en.2013-2150

Estrogens play pivotal roles in development and function of many organ systems, including the reproductive system. We have generated estrogen receptor 1 (Esr1)-knockout rats using zinc finger nuclease (ZFN) genome targeting. mRNAs encoding ZFNs targeted to exon 3 of Esr1 were microinjected into single-cell rat embryos and transferred to pseudopregnant recipients. Of 17 live births, 5 had biallelic and 1 had monoallelic Esr1 mutations. A founder with monoallelic mutations was backcrossed to a wild-type rat. Offspring possessed only wild-type Esr1 alleles or wild-type alleles and Esr1 alleles containing either 482 bp (Delta482) or 223 bp (Delta223) deletions, indicating mosaicism in the founder. These heterozygous mutants were bred for colony expansion, generation of homozygous mutants, and phenotypic characterization. The Delta482 Esr1 allele yielded altered transcript processing, including the absence of exon 3, aberrant splicing of exon 2 and 4, and a frameshift that generated premature stop codons located immediately after the codon for Thr157. ESR1 protein was not detected in homozygous Delta482 mutant uteri. ESR1 disruption affected sexually dimorphic postnatal growth patterns and serum levels of gonadotropins and sex steroid hormones. Both male and female Esr1-null rats were infertile. Esr1-null males had small testes with distended and dysplastic seminiferous tubules, whereas Esr1-null females possessed large polycystic ovaries, thread-like uteri, and poorly developed mammary glands. In addition, uteri of Esr1-null rats did not effectively respond to 17beta-estradiol treatment, further demonstrating that the Delta482 Esr1 mutation created a null allele. This rat model provides a new experimental tool for investigating the pathophysiology of estrogen action.

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CRRD Object Information
CRRD ID: 8552987
Created: 2014-05-01
Species: All species
Last Modified: 2014-05-01
Status: ACTIVE



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RGD is funded by grant HL64541 from the National Heart, Lung, and Blood Institute on behalf of the NIH.