Your browser doesn't support javascript.
loading
Establishment of L-periaxin gene knock-out RSC96 cell line / 生物工程学报
Chinese Journal of Biotechnology ; (12): 1735-1744, 2016.
Article in Chinese | WPRIM | ID: wpr-243684
ABSTRACT
Periaxin, a protein of noncompact myelin, is specifically expressed in the peripheral nervous system (PNS). There are two protein isoform L-periaxin and S-Periaxin by alternative splicing of periaxin gene, playing an important role in the initiation of myelin formation. So far, 18 different mutation sites in L-periaxin gene have been found to induce the peripheral demyelinating neurological charcot-marie-tooth diseases subtype 4F (CMT4F). The technique of activation of transcription activator-like effector nucleases (TALENS) was used to knock out the L-periaxin gene in RSC 96 cell line of Rattus. According to the design principle, the knock-out site of L-periaxin was assured to NLS domain of L-periaxin, which is target sequence of left and right arms of TALEN. The knock-out vectors of TALEN-L and TALEN-R were established and transfected into RSC96 cell. After puromycin screening, L-periaxin was knocked out successfully in RSC96 cell, which is confirmed by DNA sequence. The mutation efficiency is 21.6%. S-periaxin, not L-periaxin can be detected by Western blotting in L-periaxin gene knock-out RSC96 cell. The cell growth rate was decreased and the number of cells in G1 increased and decreased in S phase in L-periaxin gene knock-out RSC96 cell by flow cytometry and MTT assay.
Subject(s)

Full text: Available Index: WPRIM (Western Pacific) Main subject: Charcot-Marie-Tooth Disease / Cell Line / Protein Isoforms / Gene Knockout Techniques / Genetics / Membrane Proteins / Mutation Limits: Animals Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2016 Type: Article

Similar

MEDLINE

...
LILACS

LIS

Full text: Available Index: WPRIM (Western Pacific) Main subject: Charcot-Marie-Tooth Disease / Cell Line / Protein Isoforms / Gene Knockout Techniques / Genetics / Membrane Proteins / Mutation Limits: Animals Language: Chinese Journal: Chinese Journal of Biotechnology Year: 2016 Type: Article