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Because these novel compounds will be selected in vitro, it is crucial to extensively validate in vivo the selective toxicity of these drugs toward CSCs, utilizing primary tumor xenografts as a preclinical step.
Artificial ribozymes can be selected in vitro to catalyze diverse chemical reactions [1], [2], [3], [4].
In an alternative approach, recombinant single-chain Fv (scFv) antibodies, which preserve the binding specificities of monoclonal antibodies within the framework of a single small polypeptide, can be selected in vitro and expressed intracellularly as "intrabodies" to probe huntingtin and α-synuclein in living cells [18], [19].
Specific protein binders can thus be selected in vitro and used to reprogram developmental systems in vivo directly at the protein level, thereby bypassing some limitations of approaches that function at the DNA or the RNA level.
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Compared to antibodies, aptamers have several advantages, as they are selected in vitro which also enables selection against toxic or weakly immunogenic targets.
For experimental evolution, new artificial RNA aptamers and RNA enzymes (ribozymes) have been selected in vitro.
A random peptide library was generated for fusion to the N-terminal of EGF, and polypeptides exhibiting affinity were selected in vitro by ribosome display.
Several aptamer RNAs have been selected in vitro that bind to otherwise weakly fluorescent small molecules and enhance their fluorescence several thousand-fold.
Nucleic acid aptamers are selected in vitro from single stranded DNA or RNA ligands containing random sequences of up to a few hundred nucleotides.
Aptamers are short single-stranded nucleic acid molecules that are selected in vitro from a large random sequence library based on their high and specific affinity to a target molecule by a process known as SELEX.
Aptamers that are selected in vitro from random pools of DNA or RNA molecules by SELEX (Systematic evolution of ligands by exponential enrichment) technique have been extensively explored for analytical and biomedical applications.
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