PT67 cell line

The RetroPack PT67 Cell Line (Cat. No. 631510) is the recommended packaging cell line for creating a stable producer cell line for continuous production of retrovirus. This retroviral packaging system includes an NIH/3T3-based packaging cell line that expresses the 10A1 viral envelope. Virus packaged

using RetroPack PT67 cells can be used to infect a broad range of mammalian cells, including human,

mouse and rat cell types. The virus is able to enter cells via two different surface molecules, the

amphotropic retrovirus receptor (Ram-1) and the GALV receptor. NOTE: If you wish to make retrovirus via transient transfection, which is the preferred method for most researchers, we recommend the Retro-X Universal Packaging System, or the AmphoPack 293 or

EcoPack 2-293 Cell Lines.

Retro-X Retroviral Packaging Cell Lines To produce recombinant retrovirus for target cell infection, retroviral transfer vectors must be transfected into retroviral packaging cells that provide the gag, pol, and env genes in trans. Various options are available, and

all retroviral packaging cell lines and systems are compatible with all retroviral transfer vectors sold by Clontech, including MMLV- and MSCV-based retroviruses. Table 1 summarizes the retroviral packaging cell lines available from Clontech and Table 2 lists the host range of retroviruses created with the different types of commonly used envelopes. ? The Retro-X Universal Packaging System (Cat. No. 631530) is our premium retroviral packaging system featuring the GP2-293 packaging cell line. All four commonly used envelopes are supplied on

separate vectors that you cotransfect with your transfer vector. Envelope choices include VSV-G, eco,

ampho, and 10A1 to allow you to choose the tropism that is most appropriate for your target cells (see Table 2). ? The EcoPack? 2-293 Cell Line (Cat. No. 631507) is an ecotropic HEK 293-based packaging cell line that is also easy to transfect and produces retrovirus with a tropism that limits infection to mouse and rat cells*. The viral envelope protein is integrated and expressed by EcoPack 2-293, and

recognizes the ecotropic receptor (mCAT1) found only on the surface of these cell types. *NOTE: It is possible to transduce human cells with ecotropic retrovirus provided that the cells are

pretreated with Ecotropic Receptor Booster (Cat. No. 631471; Section III.J). Ecotropic Receptor Booster consists of a vial of concentrated exosome-like vesicles that are densely coated with the

mCAT-1 receptor protein. When the booster vesicles are applied to your cells, they fuse with the

plasma membrane, resulting in an increased level of the receptor on the cell surface.

? The AmphoPack-293 Cell Line (Cat. No. 631505) can be used to produce recombinant MMLV-based

viral particles that infect a broad range of mammalian cells, including human, mouse, and rat cells.

AmphoPack 293 is also derived from an HEK 293-based cell line that is easily transfected to produce high viral titers. The viral envelope protein recognizes the amphotropic receptor (Ram-1).

? The Pantropic Retroviral Expression System (Cat. No. 631512) also features the GP2-293

packaging cell line, which expresses gag and pol proteins. The envelope protein is supplied on a

separate plasmid, pVSV-G, which is cotransfected with your transfer vector. Delivery using a VSV-G envelope is not dependent on a cell surface receptor; instead, it mediates viral entry through lipid

binding and plasma membrane fusion. Two transfer expression vectors are included with the Pantropic

System, giving you the choice of using a retroviral 5' LTR or the Drosophila hsp70 promoter to

express your gene of interest.

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