De novo production of new knockout mouse lines for the next phase of the KOMP2 Project

为KOMP2项目下一阶段重新生产新的基因敲除小鼠品系 UM1OD023221行政副刊 2021年8月1日-2022年7月31日 摘要 此管理补充请求目的是生成25个新的纯合子活体 基因敲除小鼠品系准备生产表型的雄性和雌性队列 即将推出的KOMP2-Phase3项目。为了做到这一点，这一补充将为生产活动提供资金。 在三个具体目标下。具体目标1涉及基因选择和外显性KO等位基因设计， CRISPR体外质量控制指南，受精卵电穿孔，胚胎移植，妊娠 以及分娩、方正(F0)基因分型、断奶和住房筛选 F0代小鼠至繁育年龄，F0代繁育及序列质量控制(QC)以确认种系 25个基因的传递(GLT)(加州大学戴维斯分校18个，TCP 7个)。具体目标2涉及N1的QC 小鼠，向GenTar报告产量，并将N2小鼠的品系扩展到繁殖年龄，这将 准备好生成用于表型分析的男性和女性队列。《特定目标3》涉及精子 为每个产生的品系冷冻保存2个杂合子雄性，并将其沉积到 加州大学戴维斯分校的MMRRC。这项工作将于2022年7月31日前完成。

De novo production of new knockout mouse lines for the next phase of the KOMP2 Project UM1OD023221 Administrative Supplement August 1, 2021 – July 31, 2022 ABSTRACT The aim of this administrative supplement request is to generate 25 new homozygous viable gene knockout mouse lines ready for production of male and female cohorts for phenotyping in a forthcoming KOMP2-Phase3 project. To do so, this supplement will fund production activities under 3 specific aims. Specific Aim 1 involves gene selection and exdel KO allele design, CRISPR guide in vitro quality control (QC), zygote electroporation, embryo transfer, gestation and parturition, founder (F0) screening by gene specific PCR genotyping, weaning and housing F0 mice until breeding age, F0 breeding and sequence quality control (QC) to confirm germline transmission (GLT) for 25 genes (18 at UC Davis, 7 at TCP). Specific Aim 2 involves QC of N1 mice, production reporting to GenTar, and line expansion of N2 mice to breeding age which will be ready to generate male and female cohorts for phenotyping. Specific Aim 3 involves sperm cryopreservation of 2 heterozygote males for each line produced and deposition into the MMRRC at UC Davis. This work will be completed by July 31, 2022.