当前位置: X-MOL 学术ACS Synth. Biol. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Knock-Down of a ligIV Homologue Enables DNA Integration via Homologous Recombination in the Marine Diatom Phaeodactylum tricornutum.
ACS Synthetic Biology ( IF 4.7 ) Pub Date : 2018-12-21 , DOI: 10.1021/acssynbio.8b00234
Max Angstenberger 1 , Julia Krischer 1 , Ozan Aktaş 1 , Claudia Büchel 1
Affiliation  

Genetic engineering of Phaeodactylum tricornutum as a model organism for diatoms is the basis of molecular and biochemical research, and can also be used in biotechnological approaches. So far, integration of foreign DNA into the genome happens randomly by nonhomologous end joining (NHEJ), if the classical method of particle bombardment is used, with the danger of negative physiological side effects. Here we show that a putative gene for a DNA ligase IV homologue ( ligIV) in P. tricornutum codes for a functional LigIV. The knock-down of ligIV in P. tricornutum via antisense RNA drastically enhances homologous recombination (HR) by interfering with the NHEJ pathway at its central DNA ligation step done by LigIV. This enables a specific integration of DNA at desired locations, greatly enhanced transformation rates and provides a new way of specifically altering the genome of P. tricornutum.

中文翻译:

敲除ligIV同源物可通过同源重组在海洋硅藻Phaeodactylum tricornutum中实现DNA整合。

作为角质层硅藻的典范生物的三角角拟杆菌(Phaeodactylum tricornutum)的基因工程是分子和生化研究的基础,也可以用于生物技术方法中。到目前为止,如果使用经典的粒子轰击方法,则通过非同源末端连接(NHEJ)随机将外源DNA整合到基因组中,这会带来负面的生理副作用。在这里,我们显示了在角果疟原虫中DNA连接酶IV同源物(ligIV)的推定基因编码功能性LigIV。通过反义RNA敲除三角叶青霉中ligIV的过程,通过在LigIV完成的中央DNA连接步骤中干扰NHEJ途径,从而大大增强了同源重组(HR)。这样可以在所需位置进行特定的DNA整合,
更新日期:2018-12-10
down
wechat
bug