Efficient selection for high-expression transfectants with a novel eukaryotic vector

RNA and DNA expression vectors containing genes for chloramphenicol acetyltransferase, luciferase, and beta-galactosidase were separately injected into mouse skeletal muscle in vivo. Protein expression was readily detected in all cases, and no special delivery system was required for these effects. The extent of expression from both the RNA and DNA constructs was comparable to that obtained from fibroblasts transfected in vitro under optimal conditions. In situ cytochemical staining for beta-galactosidase activity was localized to muscle cells following injection of the beta-galactosidase DNA vector. After injection of the DNA luciferase expression vector, luciferase activity was present in the muscle for at least 2 months.

https://science.sciencemag.org/content/sci/247/4949/1465.full.pdf

Direct gene transfer into mouse muscle in vivo.

Liposomal drug delivery systems: from concept to clinical applications.

Cellular uptake of the tat protein from human immunodeficiency virus

In the 10 years that have passed since the Nobel prize-winning discovery of RNA interference (RNAi), billions of dollars have been invested in the therapeutic application of gene silencing in humans. Today, there are promising data from ongoing clinical trials for the treatment of age-related macular degeneration and respiratory syncytial virus. Despite these early successes, however, the widespread use of RNAi therapeutics for disease prevention and treatment requires the development of clinically suitable, safe and effective drug delivery vehicles. Here, we provide an update on the progress of RNAi therapeutics and highlight novel synthetic materials for the encapsulation and intracellular delivery of nucleic acids.

Knocking down barriers: advances in siRNA delivery

A DNA-transfection protocol has been developed that makes use of a synthetic cationic lipid, N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA). Small unilamellar liposomes containing DOTMA interact spontaneously with DNA to form lipid-DNA complexes with 100% entrapment of the DNA, DOTMA facilitates fusion of the complex with the plasma membrane of tissue culture cells, resulting in both uptake and expression of the DNA. The technique is simple, highly reproducible, and effective for both transient and stable expression of transfected DNA. Depending upon the cell line, lipofection is from 5- to greater than 100-fold more effective than either the calcium phosphate or the DEAE-dextran transfection technique.

https://www.pnas.org/content/pnas/84/21/7413.full.pdf

Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure

This invention relates to compounds of the formula I or an optical isomer thereof wherein R1 and R2 are the same or different and are an alkyl or alkenyl group of 6 to 24 carbon atoms; R3, R4 and R5 are the same or different and are alkyl of 1 to 8 carbon atoms, aryl, aralkyl of 7 to 11 carbon atoms, or when two or three of R3, R4, and R5 are taken together to form quinuclidino, piperidino, pyrrolidino, or morpholino; n is 1 to 8; and X is a pharmaceutically acceptable anion.

N[ omega ,( omega -1)-dialkyloxy]- and N-[ omega ,( omega -1)-dialkenyloxy]-alk-1-yl-N,N,N-tetrasubstituted ammonium lipids and uses therefor

This invention relates to compounds of the formula ##STR1## or an optical isomer thereof wherein R 1 and R 2 are the same or different and are an alkyl or alkenyl group of 6 to 24 carbon atoms; R 3 , R 4 and R 5 are the same or different and are alkyl of 1 to 8 carbon atoms, aryl, aralkyl of 7 to 11 carbon atoms, or when two or three of R 3 , R 4 , and R 5 are taken together to form quinuclidino, piperidino, pyrrolidino, or morpholino; n is 1 to 8; and X is a pharmaceutically acceptable anion.

N-[ω,(ω-1)-dialkyloxy]- and N-[ω,(ω-1)-dialkenyloxy]-alk-1-yl-N,N,N,-tetrasubstituted ammonium lipids and uses therefor

This invention relates to compounds of the formula
 or an optical isomer thereof wherein R' and R 2 are the same or different and are an alkyl or alkenyl group of 6 to 22 carbons; R 3 , R 4 and R 5 are the same or different and are hydrogen, alkyl of 1 to 8 carbon atoms, aryl, aralkyl of 7 to 11 carbons, or when two or three of R 3 , R 4 , and R 5 are taken together to form quinuclidino, piperidino, pyrrolidino, or morpholino; n is 1 to 8; and X is a pharmaceutically acceptable anion.

N-(omega,omega-1-dialkoxy)- and n-(omega,omega-1-dialkenoxy)-alk-1-yl-n,n,n-trisubstituted ammonium surfactants, their preparation and pharmaceutical formulations containing them

Richard Bolton Roman

Papers

Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure

N[ omega ,( omega -1)-dialkyloxy]- and N-[ omega ,( omega -1)-dialkenyloxy]-alk-1-yl-N,N,N-tetrasubstituted ammonium lipids and uses therefor

N-[ω,(ω-1)-dialkyloxy]- and N-[ω,(ω-1)-dialkenyloxy]-alk-1-yl-N,N,N,-tetrasubstituted ammonium lipids and uses therefor

N-(omega,omega-1-dialkoxy)- and n-(omega,omega-1-dialkenoxy)-alk-1-yl-n,n,n-trisubstituted ammonium surfactants, their preparation and pharmaceutical formulations containing them

Substituted alkyl-trisubstituted ammonium surfactant