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Table 2 List of plasmids used for HEK 293T cells co-transfection

From: A CRISPR interference strategy for gene expression silencing in multiple myeloma cell lines

Name

Function

Description

pVSV-G Retroviral vector (pVSV-G)

Envelope

Expresses the G glycoprotein of the vesicular stomatitis virus, necessary for viral entry

pMDLg-pRRE (pRRE)

Packaging

Includes gag, coding for the virion main structural proteins; pol, responsible for the retrovirus-specific enzymes; and RRE, a binding site for the Rev protein

pRSV-REV (pREV)

Packaging

Expresses Rev protein, which facilitates export of the RNA from the nucleus

pLV hU6-sgRND3 #3 hUbC-dCas9-KRAB-T2a-Puro (sgRND3 #3)

Silencing RND3 expression

Expresses all machinery for CRISPRi technique: sgRNA, dCas9 and KRAB

pLV hU6-sgRND3 #4 hUbC-dCas9-KRAB-T2a-Puro (sgRND3 #4)

Silencing RND3 expression

pLV hU6-sgRNA Scramble hUbC-dCas9-KRAB-T2a-Puro (Scramble)

Transduction control