|
|
The following Cold Spring Harbor Laboratory Press protocols are brought to you by BioSupplyNet in a partnership with Sigma-Aldrich. A list of products is available to use in each protocol.
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 |
|
back to top
View all available Cold Spring Harbor Laboratory Press Protocols
Product Association Disclaimer:
The Sigma-Aldrich products listed for this specific protocol were selected either to match or to supplement the
products listed within the actual protocol. The products/reagents from Sigma-Aldrich have been qualified for
usage, but may not have been validated for this specific application. Please refer to the detailed product
description on the usage of specific products of interest.
|
