Exo+ proofreading polymerases mediate genetic analysis and its application in biomedical studies

Authors: Duan-fang LIAO, Lin-ling CHEN, Cui-ying PENG, Jia ZHANG, Kai LI


Polymerases with a proofreading function in their internal 3´ to 5´ exonuclease
possess high fidelity for DNA replication both in vivo and in vitro. The obstacle
facing Exo+ polymerases for single nucleotide polymorphism (SNP) detection could
be bypassed by using primer-3´-termini modification. This hypothesis has been
well tested using three types of modified allele specific primers with: 3´ labeling, 3´
to 5´exonuclease resistance, and 3´dehydroxylation. Accordingly, three new SNP
assaying methods have been developed to carry out genome-wide genotyping,
taking advantage of the enzymatic properties of Exo+ polymerases. These new
mutation detection assays are widely adaptable to a variety of platforms, including
multi-well plate and microarray technologies. Application of Exo+ polymerases
to genetic analysis, including genotyping that is mostly relevant to pharmacogenetics,
high-fidelity gene expression profiling, rare mutation detection and
mutation load assay, will help to accelerate the pace of personalized medicine. In
this review paper, we will first introduce three new assays that we have recently
developed, and then describe a number of their applications in pharmacogenetics
and in other biomedical studies.

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