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Moreover, the percentage of Ngn2-hENPs grafted animals that reached a grid score ≥3 was significantly higher when cells had been transduced with a non-integrating vector than with an integrating vector (Figure 1E).
OppA-KO was complemented with a single copy of His-oppAMTB or its mutants by integration into the chromosome of M. smegmatis using the integrating vector pOPP206 harboring the His-oppAMTB gene under the control of the hsp60 promoter.
Integration site selection has emerged as the major factor responsible for toxicity of an integrating vector.
Retroviral vectors are commonly employed for long-term transgene expression via integrating vector technology.
Thus, these studies have defined an upper limit of viral transduction and thereby help define the therapeutic index of integrating vector systems for various gene therapy strategies.
An additional integrating vector was developed that confers more robust kanamycin resistance when integrated at single copy into the chromosome.
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Because its mechanism of integration requires DNA sequence recognition, φC31 integrase has a more restricted integration profile than other integrating vectors such as retroviruses and transposons [11].
Notably SV1 does not encode a homologue to SCO4383, or fragments of SCO4383 that could compensate for its truncation by integration of SV1 derived integrating vectors.
We therefore tested whether the integration frequencies of ϕC31 or ϕBT1 derived integrating vectors were affected if the recipient already contained pBF3 integrated at the SV1 attB site.
In the future, site-directed integration might eventually substitute for (semi-) randomly integrating vectors, provided comparable gene delivery and recombination efficiencies can be achieved.
These vectors allow for transient expression of exogenous genes without integration into the host genome, although with a low efficiency compared to integrating vectors [5].
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