Novel therapies for cancer treatment : designing high affinity selective ligands against SIRT1 enzyme
type: CreativeWork
description: The word ‘Sirtuin’ or Sir2 proteins are a class of proteins that possess either mono-ADP-ribosyltransferase, or deacylase activity, including deacetylase. SIRT1 is the most studied mammalian Sirtuins and predominantly localised in the nucleus and cytoplasm. Many Sirtuins targets are involved in cancer and in many types of cancers, SIRT1 is found to be overexpressed. Recent observations support SIRT1 being both an oncogene and a tumour suppressor, depending on the cancer etiology and type of tissue. To answer the question “How can SIRT1 behave as a tumour suppressor?”, highly selective ligands (aptamers) were developed against SIRT1 enzyme as the first step towards the development of an alternative chemotherapy for cancer diseases. The objectives of current study are to: (i) produce by in vitro SELEX procedures, SIRT1 binding single-stranded DNA (ssDNA) aptamers; (ii) characterise the interactions between selected aptamers and SIRT1 in vitro and determine their equilibrium dissociation constant (KD) values and; (iii) investigate the effects of selected aptamers on cancer cell lines. To achieve these objectives, ssDNA aptamers capable of binding SIRT1 enzyme were generated in vitro using a sequential approach known as SELEX. A total of eight novel SIRT1 aptamers (circular and linear), four circular aptamers from 8 rounds of circularisation-SELEX procedure, and the other four linear aptamers from 12 rounds of a basic-SELEX procedure were generated.The initial screening using the Fluor de Lys-SIRT1 assay for SIRT1 enzymatic activity in vitro indicated that an activator SIRT1 enzyme (circular3, circular4, linear3 and linear4) were obtained, these aptamers showed acceptable values of Km and Vmax to SIRT1 enzyme in kinetic characterisation studies. After equilibrium binding characterisation study of both linear and circular aptamers by SPR, it was show that circular3 and linear3 aptamers are good binder to SIRT1 enzyme, with the low KD constant (27.07± 0.959 nM and 48.3± 0…
abstract: The word ‘Sirtuin’ or Sir2 proteins are a class of proteins that possess either mono-ADP-ribosyltransferase, or deacylase activity, including deacetylase. SIRT1 is the most studied mammalian Sirtuins and predominantly localised in the nucleus and cytoplasm. Many Sirtuins targets are involved in cancer and in many types of cancers, SIRT1 is found to be overexpressed. Recent observations support SIRT1 being both an oncogene and a tumour suppressor, depending on the cancer etiology and type of tissue. To answer the question “How can SIRT1 behave as a tumour suppressor?”, highly selective ligands (aptamers) were developed against SIRT1 enzyme as the first step towards the development of an alternative chemotherapy for cancer diseases. The objectives of current study are to: (i) produce by in vitro SELEX procedures, SIRT1 binding single-stranded DNA (ssDNA) aptamers; (ii) characterise the interactions between selected aptamers and SIRT1 in vitro and determine their equilibrium dissociation constant (KD) values and; (iii) investigate the effects of selected aptamers on cancer cell lines. To achieve these objectives, ssDNA aptamers capable of binding SIRT1 enzyme were generated in vitro using a sequential approach known as SELEX. A total of eight novel SIRT1 aptamers (circular and linear), four circular aptamers from 8 rounds of circularisation-SELEX procedure, and the other four linear aptamers from 12 rounds of a basic-SELEX procedure were generated.The initial screening using the Fluor de Lys-SIRT1 assay for SIRT1 enzymatic activity in vitro indicated that an activator SIRT1 enzyme (circular3, circular4, linear3 and linear4) were obtained, these aptamers showed acceptable values of Km and Vmax to SIRT1 enzyme in kinetic characterisation studies. After equilibrium binding characterisation study of both linear and circular aptamers by SPR, it was show that circular3 and linear3 aptamers are good binder to SIRT1 enzyme, with the low KD constant (27.07± 0.959 nM and 48.3± 0…
author: Al-Sudani, BT
language: en
date published: 2017
date modified: 2026-07-24T04:26:35Z
provider: U. of Salford
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: U. of Salford