• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Gene Therapy and Molecular Biology >Coding end resolution in scid recombination-induciblecell lines
【24h】

Coding end resolution in scid recombination-induciblecell lines

机译：Scid重组诱导细胞系的编码末端分辨率

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

VDJ recombination is the mechanism by which antigen receptor genes are assembled. The site-specific cleavage mediated by recombination activating gene (RAG1 and RAG2) proteins generates two types of broken DNA ends: blunt signal ends and hairpin coding ends. The standard joining of these ends to form signal joints and coding joints employs several proteins involved in double strand break (DSB) repair, including KU70/80, the catalytic subunit of DNA- dependent protein kinase (DNA-PKcs), XRCC4 and ligase IV. The cells from severe combined immunodeficient (scid) mice are defective in resolving recombination coding ends due to a point mutation in the DNA-PKcs gene. To study the effect of the scid defect on coding end resolution, we have established recombination-inducible cell lines from scid mice. These cells, at the nonpermissive temperature, actively initiate recombination at the endogenous light chain loci and produce large amounts of hairpin coding ends. After returning to the permissive temperature, scid cells are capable of resolving these coding ends. However, unlike the coding end resolution in normal cells, which is a rapid and regulated process, the resolution of hairpin coding ends in scid cells is slow and error prone. The resulting coding joints contain extensive nucleotide deletions. In addition, the interlocus recombination products are found at much higher frequency in scid cells than in their normal cell counterparts. Our results suggest that functional DNA-PKcs may play an important role in facilitating effective V(D)J recombination and minimizing chromosomal instability.
机译：VDJ重组是组装抗原受体基因的机制。重组激活基因（RAG1和RAG2）蛋白介导的位点特异性切割产生两种类型的DNA断裂末端：钝信号末端和发夹编码末端。这些末端的标准连接形成信号接头和编码接头，采用了几种参与双链断裂（DSB）修复的蛋白质，包括KU70 / 80，DNA依赖性蛋白激酶（DNA-PKcs）的催化亚基，XRCC4和连接酶IV。 。由于DNA-PKcs基因中的点突变，来自严重的合并免疫缺陷（scid）小鼠的细胞在解析重组编码末端方面存在缺陷。为了研究scid缺陷对编码末端分辨率的影响，我们建立了来自scid小鼠的重组诱导细胞系。这些细胞在不允许的温度下，主动在内源轻链基因座上启动重组，并产生大量的发夹编码末端。恢复到允许的温度后，scid细胞能够解析这些编码末端。但是，与正常细胞中的编码末端分辨率不同（这是一个快速且受调控的过程），scid细胞中发夹编码末端的分辨率较慢且容易出错。所得的编码接头包含广泛的核苷酸缺失。另外，在scid细胞中发现间质重组产物的频率比在其正常细胞对应物中高得多。我们的结果表明，功能性DNA-PKcs可能在促进有效的V（D）J重组和最小化染色体不稳定中起重要作用。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号