ENHANCED PROTEIN TRANSDUCTION
Methods of enhanced protein transduction are provided. Aspects of the methods include contacting a cell with a transduction protein, where the transduction protein includes both a protein-of-interest domain and a protein transduction domain, and a nucleic acid transfection agent. Also provided are systems and kits that find use in practicing methods according to embodiments of the invention. The methods, systems and kits find use in a variety of different applications.
1 . A method of transducing a cell with a protein, the method comprising contacting the cell with:
(a) a transduction protein comprising:
(i) a protein-of-interest (POI) domain; and
(ii) a protein transduction domain (PTD); and
(b) a nucleic acid transfection agent;
under conditions sufficient to transduce the cell with the protein.
2 . The method according to claim 1 , wherein the transduction protein comprises a distinct PTD.
3 . The method according to claim 2 , wherein the distinct PTD comprises from 5 to 100 amino acids.
4 . The method according to claim 3 , wherein the distinct PTD is selected from the group consisting of PTD found in TAT, VP22,antp and poly-arginine.
5 . The method according to claim 1 , wherein the protein comprises a distributed PTD.
6 . The method according to claim 5 , wherein the distributed PTD comprises a basic patch made up of 3 or more non-sequential basic amino acid residues.
7 . The method according to claim 6 , wherein the number percentage of basic amino acid residues in the basic patch is 30 or more.
8 . The method according to claim 1 , wherein the nucleic acid transfection reagent is selected from the group consisting of Xfect™ transfection reagent, Lipofectamine™ LTX transfection reagent, Lipofectamine 2000™ transfection reagent, SiQuest™ transfection reagent, Transit-siQuest™ transfection reagent, Transit-TKO™ transfection reagent, Transit-LTI™ transfection reagent, Transit-Jurkat™ transfection reagent, Transit-2020™ transfection reagent; chloroquine, PEG, and combinations thereof.
9 . The method according to claim 1 , wherein the method further comprises evaluating the cell to determine whether the cell has been transduced with the transduction protein.
10 . The method according to claim 1 , wherein the method is an in vitro method.
11 . A transduction system comprising:
(a) a host cell;
(b) a transduction protein comprising:
(i) a POI domain; and
(ii) a PTD; and
(c) a nucleic acid transfection agent.
12 . The system according to claim 11 , wherein the transduction protein comprises a distinct PTD.
13 . The system according to claim 12 , wherein the distinct PTD comprises from 5 to 100 amino acids.
14 . The system according to claim 13 , wherein the distinct PTD is selected from the group consisting of PTD found in TAT, VP22, antp and poly-arginine.
15 . The system according to claim 11 , wherein the transduction protein comprises a distributed PTD.
16 . The system according to claim 15 , wherein the distributed PTD comprises a basic patch made up of 3 or more non-sequential basic amino acid residues.
17 . The system according to claim 16 , wherein the number percentage of basic amino acid residues in the basic patch is 30 or more.
18 . The system according to claim 11 , wherein the nucleic acid transfection reagent is selected from the group consisting of from the group consisting of Xfect™ transfection reagent, Lipofectamine™ LTX transfection reagent, Lipofectamine 2000™ transfection reagent, SiQuest™ transfection reagent, Transit-siQuest™ transfection reagent, Transit-TKO™ transfection reagent, Transit-LTI™ transfection reagent, Transit-Jurkat™ transfection reagent, Transit-2020™ transfection reagent; chloroquine, PEG, and combinations thereof.
19 . A kit comprising:
(a) at least one of:
(i) a vector comprising an expression cassette comprising a distinct PTD encoding element; and a cloning site; and
(ii) a transduction protein; and
(b) a nucleic acid transfection agent.
20 . The kit according to claim 19 , wherein the nucleic acid transfection reagent is selected from the group consisting of from the group consisting of Xfect™ transfection reagent, Lipofectamine™ LTX transfection reagent, Lipofectamine 2000™ transfection reagent, SiQuest™ transfection reagent, Transit-siQuest™ transfection reagent, Transit-TKO™ transfection reagent, Transit-LTI™ transfection reagent, Transit-Jurkat™ transfection reagent, Transit-2020™ transfection reagent; chloroquine, PEG, and combinations thereof.