Knock-down by transfection of shRNA expression vector by electroporation
Knock-down by transfection of shRNA expression vector by electroporation
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NEPA21 / CUY21 Applications |
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[In Ovo] Knock-down by transfection of shRNA expression vector by electroporation
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(A) shRNA expression vector is electroporated as shown above. (B) Select target DNA sequence of 19 to 21 mer. Sense and antisense sequence were linked to a nucleotide spacer as a loop and put into expression vector that is driven by U6 or H1 promoter. Commercially available expression vector that dreives expression by mouse U6 promoter is effective in chick embryos. (C) After transcription, hairpin is digested to form siRNA, and siRNA forms RISK (RNA-induced silencing complex) to degrade the target mRNA. . |
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24 hours after electroporation. (A) Transfection is monitored by co-electroporated GFP fluorescence. (B), (C) Application of siRNA against En2 by electroporation shows degradation of En2 mRNA (B). (C) Control side . |
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Graduate School of Life Sciences, and Institute of Development, Aging and Cancer, Tohoku University Harukazu Nakamura and Tatsuya Katahira *Mechanisms of Development, Volume 121, Issue 9, Pages 1137-1143, September 2004 *Development Growth & Differentiation, Volume 45, Issue 4, Pages 361-367, August 2003 . |
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