Micro-minicircle gene therapy: Implications of size on fermentation, complexation, shearing resistance, and expression

Sökning: L773:2162 2531 > Micro-minicircle ge...

Micro-minicircle gene therapy : Implications of size on fermentation, complexation, shearing resistance, and expression

QC 20140414
A3337
The minicircle (MC), composed of eukaryotic sequences only, is an interesting approach to increase the safety and efficiency of plasmid-based vectors for gene therapy. In this paper, we investigate micro-MC (miMC) vectors encoding small regulatory RNA. We use a construct encoding a splice-correcting U7 small nuclear RNA, which results in a vector of 650 base pairs (bp), as compared to a conventional 3600 bp plasmid carrying the same expression cassette. Furthermore, we construct miMCs of varying sizes carrying different number of these cassettes. This allows us to evaluate how size influences production, supercoiling, stability and efficiency of the vector. We characterize coiling morphology by atomic force microscopy and measure the resistance to shearing forces caused by an injector device, the Biojector. We compare the behavior of miMCs and plasmids in vitro using lipofection and electroporation, as well as in vivo in mice. We here show that when the size of the miMC is reduced, the formation of dimers and trimers increases. There seems to be a lower size limit for efficient expression. We demonstrate that miMCs are more robust than plasmids when exposed to shearing forces, and that they show extended expression in vivo.

Wiklander, O. P. B.Karolinska Institutet,Karolinska Institute, Sweden (författare)
Badal-Tejedor, M.RISE,SP Kemi Material och Ytor (författare)
Turunen, J.Karolinska Institutet,Karolinska Institute, Sweden (författare)
Nordin, J. Z.Karolinska Institutet,Karolinska Institute, Sweden (författare)
Hallengärd, D.Karolinska Institutet,Karolinska Institute, Sweden (författare)
Wahren, B.Karolinska Institutet,Karolinska Institute, Sweden (författare)
Andaloussi, S. E. L.Karolinska Institute, Sweden; University of Oxford, United Kingdom (författare)
Rutland, Mark W.KTH,Yt- och korrosionsvetenskap,KTH Royal Institute of Technology, Sweden(Swepub:kth)u1ml9bxf (författare)
Smith, C. I. E.Karolinska Institutet,Karolinska Institute, Sweden; CI Edvard Smith MCG group, Sweden (författare)
Lundin, K. E.Karolinska Institutet,Karolinska Institute, Sweden (författare)
Blomberg, P. (författare)
Karolinska InstitutetKarolinska Institute, Sweden (creator_code:org_t)

Av författaren/redakt...: Stenler, S.; Wiklander, O. P. ...; Badal-Tejedor, M ...; Turunen, J.; Nordin, J. Z.; Hallengärd, D.; visa fler...; Wahren, B.; Andaloussi, S. E ...; Rutland, Mark W.; Smith, C. I. E.; Lundin, K. E.; Blomberg, P.; visa färre...

Om ämnet

MEDICIN OCH HÄLSOVETENSKAP: MEDICIN OCH HÄLS ...; och Medicinsk biotek ...; och Medicinsk biotek ...

Kungliga biblioteket hanterar dina personuppgifter i enlighet med EU:s dataskyddsförordning (2018), GDPR. Läs mer om hur det funkar här.
Så här hanterar KB dina uppgifter vid användning av denna tjänst.