Sprague Dawley rat Tff2-/- rat
| Status | Available to order |
| EMMA ID | EM:13599 |
| International strain name | Sprague Dawley rat Tff2-/- rat |
| Alternative name | Sprague Dawley rat Tff2-/- rat |
| Strain type | Endonuclease-mediated |
| Allele/Transgene symbol | Tff2 (NC 005119) |
| Gene/Transgene symbol | Tff2 |
Information from provider
| Provider | Peter Taylor |
| Provider affiliation | School of Pharmacy, University College London |
| Genetic information | CRISPR/Cas9 generated knockout in Sprague-Dawley rat |
| Phenotypic information | Homozygous:Increased susceptibility of neonates (2-5 postpartum) to colonisation and infection. Otherwise no obvious phenotypic characteristicsHeterozygous:Appear and behave as normal |
| Breeding history | Two pairs of single-guide (sg) RNAs were designed to cleave together in order to generate a ~1.8kb deletion (4894 bp) between the target sites, removing the gene encoding Tff2 from the NC 005119 region of the genome. The most active sgRNAs were selected in cultured eukaryotic cells; the design and synthesis of donor DNA containing the desired mutation were facilitated by Horizon in-house bioinformatic programs and were validated by sequencing. Donor DNA along with Cas9/sgRNA reagent was delivered by microinjection into fertilized embryos to create the desired mutation; a maximum of two microinjection sessions were performed. The most potent sgRNA with minimal off-target potential was assembled into a ribonucleoprotein complex with Cas9 endonuclease and, together with the donor DNA, was delivered into zygotes from Sprague-Dawley rats followed by embryo transfer into pseudo-pregnant females. For gestation and identification of the founder mutant phase, tissue biopsies were obtained at approximately two weeks postpartum. Viable progeny was analyzed for the presence of the desired mutation by genomic PCR and DNA sequencing. Fifteen animals were screened and four founders contained a 1.8kb deletion; two rats were bred to the F1 heterozygous stage with wildtype Sprague-Dawley rats and nineteen F1 heterozygotes were identified as carrying the Tff2 mutation by PCR-mediated genotyping and DNA sequence analysis. Loss of Tff2 did not impact on the wellbeing of Tff2-/- animals, at least during the first fourteen weeks of life: pups remained healthy and homozygous adults bred in comparable fashion to wildtype. |
| References |
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| Homozygous fertile | not known |
| Homozygous viable | yes |
| Homozygous matings required | no |
| Immunocompromised | not known |
Information from EMMA
| Archiving centre | Mary Lyon Centre at MRC Harwell, Oxford, United Kingdom |
Literature references
- Loss of Trefoil Factor 2 Sensitizes Rat Pups to Systemic Infection with the Neonatal Pathogen Escherichia coli K1.;McCarthy Alex J, Birchenough George M H, Taylor Peter W, ;2019;Infection and immunity;87;1438-42; 30833331
- [Frequency of psychiatric disorders in nonpsychiatric hospital patients].;Künsebeck H W, Lempa W, Freyberger H, ;1984;Deutsche medizinische Wochenschrift (1946);109;; 6479038