Silencing the HSL Gene in Human 293 CellsPublic
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Hormone-sensitive lipase (HSL) is thought to be the major enzyme responsible for the release of free fatty acids from adipose tissues. HSL hydrolyses triacylglycerides to diacylglyceride, releasing free fatty acids from fats (lipolysis). Using RNAi to silence the gene would be advantageous in the study of obesity, so three highly effective 19-nucleotide short hairpin RNAs (shRNAs 1-3) were designed against the HSL gene. Their ability to reduce HSL protein levels was verified in human 293 cells using immunoblots. shRNA-2 sequence was inserted into an adenovirus genome, and a high titer viral solution was created which could be used in the future to silence the HSL gene in vivo.
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