However, the strand placement of the desired nucleotide change and the length of nonhomologous sequences flanking target nucleotides played a crucial role in the efficiency of chimeraplast-mediated gene correction.
Here, we report that efficient ZFN-mediated gene correction occurs at a transduced, transcriptionally active, mutant GFP locus by homology-directed repair, and that efficient mutagenesis by non-homologous end joining (NHEJ) occurs at the endogenous, transcriptionally silent, CCR5 locus in HEK293 Flp-In cells, using designed 3- and 4-finger ZFNs.
As expected, gRNA-3 and -4, but not gRNA-1, supported Cas9-mediated gene correction (Table 3).
Human NSC display significant proliferation potential and amenability to viral-mediated gene-correction in vitro, coupled to lack of tumorigenicity upon grafting in the CNS, where they survive, migrate and differentiate appropriately [12], [13], [14], [15], [16], also delivering neuroprotective factors [17], [18], [19], [20].
AAV mediated gene transfer with these new vectors showed long-term correction of the metabolic defect in fat-fed Ldlr−/− mice [22] and prevention of atherosclerosis in apoliporotein E deficient (ApoE−/−) mice [19] and fat-fed Ldlr−/− mice [22].
This vector has potential for targeted gene correction in hemoglobinopathies.
With its potential for protein replacement and gene correction, mod.
mRNA has tremendous potential for both gene therapy and gene correction approaches.
In this work, repair-PPRHs are shown to be a new powerful tool for gene correction.
