通过基因编辑技术构建的 p53 基因敲除小鼠表型分析及其与传统 p53 基因敲除小鼠的比较。

Phenotyping analysis of p53 knockout mice produced by gene editing and comparison with conventional p53 knockout mice.

机构信息

Department of Laboratory Animal Medicine, BK21 PLUS Program for Creative Veterinary Science Research, Research Institute for Veterinary Science and College of Veterinary Medicine, Seoul National University, Seoul, 08826, Republic of Korea.

Department of Stem Cell Biology, School of Medicine, Konkuk University, Seoul, 05029, Republic of Korea.

出版信息

Genes Genomics. 2019 Jun;41(6):701-712. doi: 10.1007/s13258-019-00785-y. Epub 2019 Apr 15.

DOI:10.1007/s13258-019-00785-y

PMID:30989490

Abstract

BACKGROUND

Knockout (KO) mice developed by homologous recombination (HR) have become useful tools to elucidate gene function. However, HR has low KO efficiency and is time-consuming, labor-intensive, and expensive. 'Gene editing' has received much attention for efficient genetic manipulation.

OBJECTIVE

As generation of KO mice is simplified, KO mice produced by HR can be feasibly reproduced using gene editing. However, phenotyping analysis and comparison between KO mice produced by these two techniques is necessary.

METHODS

We generated p53 KO mice through gene editing and compared their phenotype with the already reported HR-mediated p53 KO mice.

RESULTS

Tumors occurred in 36 (73%) of 49 homozygous KO mice and the mean age of occurrence was 23 weeks, with lymphoma (64%) and sarcoma (23%) being the most common. Tumors were also developed in 12 heterozygous mice and the mean age of occurrence was 40 weeks, with sarcoma (54%) and lymphoma (46%) in high proportion. Homozygotes had a mean life span of 157 ± 52 days and developmental abnormalities were found in females compared to in males (P < 0.05, P < 0.001).

CONCLUSION

We analyzed the basic phenotype of p53 KO mice and observed no significant difference from the conventional HR-mediated p53 KO mice.

摘要

背景

通过同源重组（HR）开发的敲除（KO）小鼠已成为阐明基因功能的有用工具。然而，HR 的 KO 效率低，耗时、费力且昂贵。“基因编辑”因其高效的遗传操作而受到广泛关注。

目的

随着 KO 小鼠的生成简化，通过 HR 产生的 KO 小鼠可以通过基因编辑来合理复制。然而，有必要对这两种技术产生的 KO 小鼠进行表型分析和比较。

方法

我们通过基因编辑生成了 p53 KO 小鼠，并将其表型与已经报道的 HR 介导的 p53 KO 小鼠进行了比较。

结果

49 只纯合 KO 小鼠中有 36 只（73%）发生了肿瘤，平均发病年龄为 23 周，最常见的是淋巴瘤（64%）和肉瘤（23%）。12 只杂合 KO 小鼠也发生了肿瘤，平均发病年龄为 40 周，肉瘤（54%）和淋巴瘤（46%）比例较高。纯合子的平均寿命为 157±52 天，与雄性相比，雌性出现发育异常（P<0.05，P<0.001）。