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Sigma-Aldrich Research Biotechnology
Lentivirus Transduction Protocol | ||
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Sigma® has prepared product lists for commonly used protocols from Cold Spring Harbor Protocols. |
View the protocol on BioSupplyNet.com.
| Selected Protocol from "Gene Transfer: Delivery and Expression of DNA and RNA", Edited by Theodore Friedmann and John Rossi. |
| Lentivirus Transduction of Hematopoietic Cells |
| Ming-Jie Li and John J. Rossi Division of Molecular Biology, Beckman Research Institute of the City of Hope, Duarte, California 91010 |
| ABSTRACT Efficient transfer and sustained expression of transgenes are among the most important issues in gene delivery technologies. The majority of hematopoietic cells, including hematopoietic stem/progenitor cells and terminally differentiated cells, such as primary T cells and macro phages, are nondividing or slowly self-renewing. These cell types are refractory to most nonviral or retroviral delivery methods. Lentiviral vectors are capable of transducing nondividing cells and maintaining long-term and sustained expression of the transgenes. Many hematopoietic cell types have been successfully transduced with lentiviral vectors carrying a variety of genes. Lentiviral vectors are becoming useful for many delivery protocols, such as long-term expression of short hairpin RNA (shRNA) and functional genetics. They may also have great potential in gene therapy. This protocol describes lentivirus-vector-based delivery of foreign genes to hematopoietic cells. The method is applicable to various cell types in experiments that require long-term transgene expression. |
| Products Available for this Protocol |
| Protocol Material Description | Product # | Product Name | Add to Cart |
| Buffers, Solutions, and Reagents | |||
| BSA | A3156 | Albumin, essentially globulin-free from bovine serum | |
| Phosphate-buffered saline (PBS) | P5368 | Phosphate buffered saline | |
| CaCl2 (2 M) | C5670 | Calcium chloride, insect cell culture tested | |
| Water | W4502 | Water | |
| Formaldehyde | F8775 | Formaldehyde solution, for molecular biology | |
| HEPES | H4034 | HEPES, Biotechnology Performance Certified | |
| NaCl | S3014 | Sodium chloride | |
| KCl | P9541 | Potassium chloride | |
| KH2PO4 | 60353 | Potassium phosphate dibasic | |
| EDTA | E7889 | Ethylenediaminetetraacetic acid disodium salt solution | |
| Na2HPO4 | S7907 | Sodium phosphate dibasic, for molecular biology | |
| Polybrene (hexadimethrine bromide) | H9268 | Hexadimethrine bromide | |
| Sodium butyrate | B5887 | Sodium butyrate | |
| Tris-HCl | T5941 | Trizma® hydrochloride, for molecular biology, Biotechnology Performance Certified, cell culture tested | |
| Dulbecco's modified Eagle's medium (DMEM) with high glucose (4500 mg/liter) | D5796 | Dulbecco's Modified Eagle's Medium - high glucose | |
| Fetal bovine serum (FBS) | F2442 | Fetal Bovine Serum, USA origin, sterile-filtered, cell culture tested, hybridoma tested | |
| Penicillin and streptomycin | P4333 | Penicillin-Streptomycin, 100 ×, liquid, stabilized, sterile-filtered, cell culture tested | |
| Iscove's modified Dulbecco's medium (IMDM) | I7633 | Iscove's Modified Dulbecco's Medium, With L-glutamine and 25 mM HEPES, without sodium bicarbonate | |
| Stem cell factor (SCF) | S7901 | Stem Cell Factor human, recombinant, expressed in Escherichia coli, powder | |
| Thrombopoietin (TPO) | T1568 | Thrombopoietin human, recombinant, expressed in Escherichia coli, lyophilized powder | |
| Cells | |||
| Cells | ECACC cell lines | Cell Lines |
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