Application of Laser Micro-irradiation for Examination of Single and Double Strand Break Repair in Mammalian Cells.
Application of Laser Micro-irradiation for Examination of Single and Double Strand Break Repair in Mammalian Cells.
复制标题
DOI：
10.3791/56265
复制
发表时间：
2017-09-05
期刊：
影响因子：
--
通讯作者：
Gassman NR
中科院分区：
文献类型：
--
作者：
Holton NW;Andrews JF;Gassman NR
存在高度协调的DNA修复途径来检测、切除和替换受损的DNA碱基，并协调DNA链断裂的修复。虽然分子生物学技术已经阐明了修复蛋白的结构，酶功能和动力学，但仍然需要了解修复是如何在细胞核内协调的。激光微辐射为诱导DNA损伤和监测修复蛋白的募集提供了强有力的工具。通过激光微辐射诱导DNA损伤可以在一定范围的波长下发生，并且用户可以在一定范围的剂量下可靠地诱导单链断裂、碱基损伤和双链断裂。在这里，激光显微照射用于检查由两种常见的共聚焦激光波长355 nm和405 nm诱导的单链和双链断裂的修复。此外，适当的表征所施加的激光剂量诱导特定的损伤混合物进行了描述，所以用户可以重复执行激光微照射数据采集和分析。
Highly coordinated DNA repair pathways exist to detect, excise and replace damaged DNA bases, and coordinate repair of DNA strand breaks. While molecular biology techniques have clarified structure, enzymatic functions, and kinetics of repair proteins, there is still a need to understand how repair is coordinated within the nucleus. Laser micro-irradiation offers a powerful tool for inducing DNA damage and monitoring the recruitment of repair proteins. Induction of DNA damage by laser micro-irradiation can occur with a range of wavelengths, and users can reliably induce single strand breaks, base lesions and double strand breaks with a range of doses. Here, laser micro-irradiation is used to examine repair of single and double strand breaks induced by two common confocal laser wavelengths, 355 nm and 405 nm. Further, proper characterization of the applied laser dose for inducing specific damage mixtures is described, so users can reproducibly perform laser micro-irradiation data acquisition and analysis.
登录
查看更多内容
影响因子：
3.8
作者：
Gassman, Natalie R.;Wilson, Samuel H.
通讯作者：
Wilson, Samuel H.
影响因子：
1
作者：
Lomax, ME;Gulston, MK;O'Neill, P
通讯作者：
O'Neill, P
影响因子：
2.8
作者：
Hanssen-Bauer, Audun;Solvang-Garten, Karin;Otterlei, Marit
通讯作者：
Otterlei, Marit
影响因子：
3.4
作者：
KRASIN, F;HUTCHINSON, F
通讯作者：
HUTCHINSON, F
DOI：
10.1073/pnas.0406048101
发表时间：
2004-09-21
影响因子：
11.1
作者：
Lan, L;Nakajima, S;Yasui, A
通讯作者：
Yasui, A