Screening of RNA molecules inhibiting human acetylcholinesterase by virtue of systematic evolution of ligands by exponential enrichment
Abstract
AIM: To acquire the specific RNA aptamers inhibiting human red blood cell (RBC) acetylcholinesterase (AChE).
METHODS: Systematic evolution of ligands by exponential enrichment (SELEX) aptamer against human red blood cell membrane AChE was selected by microtiter plate method in vitro. The specifity binding to AChE was determined by gel mobility shift analysis. Microcolorispectrophotometric method was used to measure the activity of AChE.
RESULTS: The aptamers to human RBC AChE were identified by 9 reiterative rounds. At the same concentration (2.25 micromol/L), the aptamers did not bind to the recombinant human butyrylcholinesterase (rhBChE) but specifically bound to human RBC AChE and inhibited the enzyme activity.
CONCLUSION: It is an effective way to isolate the specific AChE inhibitor from the vast oligonucleotide combinatorial library by virtue of SELEX.
Keywords:
METHODS: Systematic evolution of ligands by exponential enrichment (SELEX) aptamer against human red blood cell membrane AChE was selected by microtiter plate method in vitro. The specifity binding to AChE was determined by gel mobility shift analysis. Microcolorispectrophotometric method was used to measure the activity of AChE.
RESULTS: The aptamers to human RBC AChE were identified by 9 reiterative rounds. At the same concentration (2.25 micromol/L), the aptamers did not bind to the recombinant human butyrylcholinesterase (rhBChE) but specifically bound to human RBC AChE and inhibited the enzyme activity.
CONCLUSION: It is an effective way to isolate the specific AChE inhibitor from the vast oligonucleotide combinatorial library by virtue of SELEX.