An improved PCR strategy for fast screening of specific and random integrations in rAAV-mediated gene targeted cell clones.

ABSTRACT:

Background

Gene targeting by homologous recombination using recombinant adeno-associated virus (rAAV) is becoming a useful tool for basic research and therapeutic applications due to the remarkably high targeting frequency of rAAV virus vectors. However, the screening for the pure gene-targeted and random-integration-free primary cell clones is difficult since the cells have a limited proliferation capacity and often cannot be grown to produce sufficient DNA for non-PCR based analysis. This hampers the applications of this technology.

Findings

In this study, we have developed an improved PCR screening method, which can be used for fast screening of clones with unwanted random integration (RI) of the rAAV genome. This improved screening method includes four PCRs: a PCR for th

SUBMITTER: Luo Y 

PROVIDER: S-EPMC3154164 | biostudies-literature | 2011 Jul

REPOSITORIES: biostudies-literature