Related Experiment Video
Updated: Mar 21, 2026
Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells
Published on: July 16, 2011
[Knockout of Micro-RNA-21 Gene in DLBCL OCI-Ly3 cells by TALEN Technique]
Bu-Yuan Chen1,2, Jian-Da Hu1, Ren-Zhang Lin2
1Fujian Medical University Union Hosspital, Fujian Institute of Hematology, Fujian Provincal Key Laboratory of Hematologic Diseases, Fuzhou 350001, Fujian Province, China.
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.
Conclusion:
This strategy may be applied to knockout any microRNA of interest even in hemapoietic cell lines with very low transfection efficiency.
