All experiments, excluding those using neonatal mice, utilized 6–8-wk-old male MyD88 knockout (KO) mice (gift of Jian-Dong Li, Georgia State University, Atlanta, GA, USA), IL-18 KO mice (Jackson Laboratories), IL-1R KO (Jackson Laboratories), NLRP3 KO mice (Jackson Laboratories, Bar Harbor, ME, USA) or C57BL/6 offspring from C57BL/6 mice originally purchased from Jackson Laboratories. Experiments involving neonatal mice used 6-d-old neonates bred in-house.
Nlrp3 ko mice
Manufactured by Jackson ImmunoResearch
Sourced in United States
NLRP3 KO mice are genetically modified mice that have had the NLRP3 gene knocked out. The NLRP3 gene is responsible for the production of the NLRP3 protein, which is a key component of the NLRP3 inflammasome. These mice can be used in research to study the role of the NLRP3 inflammasome in various biological processes and disease models.
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4 protocols using nlrp3 ko mice
1
Immune Pathways in Mouse Models
2
Acclimation and Handling of NLRP3 KO Mice
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Male NLRP3 KO mice and wild-type mice (8 weeks old) with C57BL/6 background were purchased from the Jackson Laboratory, experimental Animal Center (Shanghai, China). After arrival, the mice were housed in a temperature and humidity-controlled animal experiment laboratory with a 12-h light/dark cycle (lights on at 7:00 a.m.), and standard laboratory water and chow were provided randomly. To reduce environmental effects, those mice were allowed to adapt to laboratory conditions for 2 weeks before use.
The experimental procedures involving animals were performed by the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines (Percie du Sert et al., 2020 (link); Karp et al., 2015 (link)), and approved by the Animal Experimental Ethical Inspection Form of Southeast University.
The experimental procedures involving animals were performed by the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines (Percie du Sert et al., 2020 (link); Karp et al., 2015 (link)), and approved by the Animal Experimental Ethical Inspection Form of Southeast University.
3
Genetic Manipulation of Mice for Kidney Damage Research
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Agtr1a KO and NLRP3 KO mice on the C57BL/6 background were purchased from Jackson laboratory and were backcrossed to the 129/SvEv mouse strain for ≥6 generations to increase their susceptibility to kidney damage[26 (link)]. Thereafter, heterozygotes on the 129/SvEv background were intercrossed to yield knock-out (KO) and wild-type (WT) littermates for our experiments. Eight- to 12-week-old male mice were used for the experiments. All experiments described in this manuscript were included in an animal use protocol approved by the Ethics Review Committees for Animal Experimentation of Southeast University.
4
Genetically Modified Mouse Models for Inflammation
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Mice were housed in a 12:12 h light–dark cycle with food and water ad libitum. For all experiments, C57B/6 mice of the same age were used. For the IL1β bioassay, we used mice in which the entire coding sequence of Nlrp3 was replaced with a Neo cassette (Nlrp3 KO mice, The Jackson Laboratory, Bar Harbor, ME USA; #B6.129S6- Nlrp3tm1Bhk/J). Genotyping was performed with 5x HOT FIREPol EvaGreen® qPCR Mix and a total volume of 20 µL and primer sequences as follows: mutant forward 5′-TGCCTGCTCTTTACTGAAGG-3′, wild type forward 5′-TCAGTTTCCTTGGCTACCAGA-3′, and common reverse 5′-TTCCATTACAGTCACTCCAGATGT-3′, as described by The Jackson Laboratory. B6.129P2-Lyz2tm1(cre)Ifo/J mice (The Jackson Laboratory) were crossed with C57B/6 mice bearing LoxP sites upstream and downstream of Tsc22d3 exon 6 to obtain myeloid-specific Gilz knockout (KO) mice. Breeding and genotyping were performed as described previously [47 (link)].
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