title: [Knockout of Micro-RNA-21 Gene in DLBCL OCI-Ly3 cells by TALEN Technique].
abstract: OBJECTIVE
  To explore an efficient way to knockout microRNA genes in hemapoietic cell lines with a very low transfection efficiency, so as to facilitate the study of microRNA function in hematopoietic malignancies.
  
  
  METHODS
  TALE-nucleases was utilized to knockout the microRNA-21 gene in human diffuse large B-cell lymphoma cells (OCI-Ly3). The OCI-Ly3 single cell clones without expression of miR-21 were established through eGFP(+) enrichment, PCR screening, and microRNA quantification. Finally, the miR-21 changes of mutant clones were identified by sequencing.
  
  
  RESULTS
  Four miR-21-knockouted OCI-Ly3 single-cell-derived clones were established after 2 round transfection and screening. The miR-21 knockout efficiency was around 10/10(6) original cells. Sequencing the mutant clones indicated that miR-21 expression could be drastically reduced by simply altering sequences immediately adjacent to the microRNA duplex.
  
  
  CONCLUSIONS
  This strategy may be applied to knockout any microRNA of interest even in hemapoietic cell lines with very low transfection efficiency.
authors.affiliation: Fujian Medical University Union Hosspital, Fujian Institute of Hematology, Fujian Provincal Key Laboratory of Hematologic Diseases, Fuzhou 350001, Fujian Province, China.|Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope Duarte, CA, USA.
authors.forename: Bu-Yuan
authors.initials: BY
authors.lastname: Chen
authors.affiliation: Fujian Medical University Union Hosspital, Fujian Institute of Hematology, Fujian Provincal Key Laboratory of Hematologic Diseases, Fuzhou 350001, Fujian Province, China.
authors.forename: Jian-Da
authors.initials: JD
authors.lastname: Hu
authors.affiliation: Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope Duarte, CA, USA. E-mail: RLin@coh.org.
authors.forename: Ren-Zhang
authors.initials: RZ
authors.lastname: Lin
authors.affiliation: Fujian Medical University Union Hosspital, Fujian Institute of Hematology, Fujian Provincal Key Laboratory of Hematologic Diseases, Fuzhou 350001, Fujian Province, China.|Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope Duarte, CA, USA. E-mail: chenxinji@sina.com.
authors.forename: Xin-Ji
authors.initials: XJ
authors.lastname: Chen
mesh_terms: D045744:Cell Line, Tumor; D055786:Gene Knockout Techniques; D006801:Humans; D016403:Lymphoma, Large B-Cell, Diffuse / Q000235:genetics*; D035683:MicroRNAs / Q000235:genetics*; D014162:Transfection
chemical_list: C524940:MIRN21 microRNA, human; D035683:MicroRNAs