Methods for gene delivery in large animal models

10 posts / 0 new
Last post
Scot E. DOwd Ph.D.
Scot E. DOwd Ph.D.'s picture
Methods for gene delivery in large animal models

Anybody got ideas? Good topic

Soudabeh's picture
Scot E. DOwd Ph.D. wrote

Scot E. DOwd Ph.D. wrote:

Anybody got ideas? Good topic

Methods such as: Electroporation is used in order to eliminate compolications associated with viral gene delivery system. Also liver targeted gene therapy was done in canine.

jangy's picture
LOL, we tried electroporation

LOL, we tried electroporation AND liver targetted!!

scolix's picture
HIV-1-Tat-derived cell

HIV-1-Tat-derived cell-penetrating peptide (CPP) provides a means to deliver fusion proteins into the brain.

Viral mediated gene transfer is great. Using AAV is harmless.

nin1318's picture
i have found that UPS, fed-ex

i have found that UPS, fed-ex and priority mail all do a wonderful delivery job. sometimes when time is really an issue and i need to get a grant delivered to NIH or something...i'll go with a local courrier for super-fast service...


sorry about that i am feeling like a smart-alec. :-)

Fraser Moss
Fraser Moss's picture
I think Biolistics gets used

I think Biolistics gets used for this purpose - "The Helios Gene Gun"

Basically it works by blasting gold particles coated with your DNA of interest into the organ or region you want to transfect.

Jon Moulton
Jon Moulton's picture
For knocking down a gene,

For knocking down a gene, there have been some promising reports using uncharged antisense oligos and arginine rich peptides to bring the oligos across cell membranes. However, this won't insert a gene. Richard Kinney of the US CDC presented some preliminary mouse data at the 2005 ASV meeting, but nothing showing efficacy of these peptide conjugates has been published in the journals yet.

Nelson MH, Stein DA, Kroeker AD, Hatlevig SA, Iversen PL, Moulton HM. Arginine-rich Peptide conjugation to morpholino oligomers: effects on antisense activity and specificity. Bioconjug Chem. 2005 Jul-Aug;16(4):959-66.

Moulton HM, Nelson MH, Hatlevig SA, Reddy MT, Iversen PL. Cellular uptake of antisense morpholino oligomers conjugated to arginine-rich peptides. Bioconjug Chem. 2004 Mar-Apr;15(2):290-9.

Moulton HM, Hase MC, Smith KM, Iversen PL. HIV Tat peptide enhances cellular delivery of antisense morpholino oligomers. Antisense Nucleic Acid Drug Dev. 2003 Feb;13(1):31-43.

- Jon

Braisler's picture
A better and more leading

A better and more leading question would be what (organ) system are you trying to target, with what gene, for what duration? There are abundant choices, but to give an informed answer we need some more information about what you are trying to do.

Different vectors (and serotypes) have different tissue specificity. You can also think about tissue specific or inducible promoters if you want to limit expression to a particular tissue type or over a particular time frame.

atcg's picture
I don't know if this will

I don't know if this will help, but here is an exerpt from the American Society of Gene Therapy, Annual Meeting Highlights
By Roger J. Hajjar, MD, Member, Cardiovascular Committee Member

"Dr. Roger Hajjar reviewed the techniques for gene delivery in rodents and large animals and his groups experience in targeting the sarcoplasmic reticulum calcium ATPase pump (SERCA2a) for improving calcium cycling in heart failure. Gene therapy with adeno-associated virus carrying SERCA2a is currently being considered for clinical trials in patients undergoing left ventricular assist device placement for advanced heart failure. Dr. Walter Koch reviewed approaches of intracoronary adenoviral gene delivery to the heart and showed that various agents including nitric oxide donors can improve viral infection of cardiomyocytes. Dr. Koch also showed preliminary data on the beneficial effects of the peptide inhibitor of beta receptor kinase 1 (ARK1) on ventricular function in models of heart failure. In a second session, Dr. Andrew Baker shared his experience with endothelial targeting by pre-incubating adenovirus with a bispecific antibody (Fab9B9) that binds both to the adenoviral coat and the angiotensin converting enzyme. Using this method of gene transfer, endothelial nitric oxide synthase (eNOS) induced a reduction in blood pressure in hypertensive rats. Dr. Frank Giordano has engineered zinc finger protein (ZFP) transcriptional repressors of phospholamban (which regulates SERCA2a) to improve calcium handling in heart failure. He showed that these ZFPs decrease expression of phospholamban by more than 80% resulting in accelerated calcium decay in neonatal cardiomyocytes. This highly effective and specific ZFP based approach provides an alternative to cDNA based gene therapies. Adeno-associated virus continues to gain a foothold in cardiovascular gene transfer. "

Omai's picture
Here is a very interesting

Here is a very interesting review article from the European Journal of Nature Genetics discussing large animal models and Gene therapy. It details multiple methods of gene delivery in canine, feline, equine,and bovine models from a disease specific perspective.