lentiviral vector prrlsin cppt pgk gfp wpre  (Addgene inc)

Journal: bioRxiv

Article Title: Functional Impact of CYFIP2 RNA Editing on Actin Regulation, Axon Growth, and Spinogenesis

doi: 10.1101/2025.03.04.641430

Figure Lengend Snippet: (a) IR acquisition of WB obtained from cellular lysate derived from WT Hippocampal culture (WT) and after 48 from Lentiviral transduction of shRNA targeting the 3’UTR of Cyfip2 gene (KD). Endogenous CYFIP2 protein is detected with rabbit-α-CYFIP2 antibody. In the graph CYFIP2 downregulation percentage is reported. (b) Cellular lysate of KD hippocampal cells growth for 18 DIV after lentiviral transduction with CDS of human CYFIP2 K or E variants. Exogenous CYFIP2 proteins are detected using rabbit-α-HA antibody. (c) DIV18 hippocampal cell captured by confocal microscopy. The GFP protein, which represents a marker for CYFIP2 down-regulation, is detected in green. Exogenous CYFIP2 is visible in red (rabbit-α-HA antibody -goat-α-rabbit Alexa Fluor 594). (d) CYFIP2 K/E editing levels analysed from total RNA samples obtained from wild-type hippocampal cell (WT) or CYFIP2 knock-down hippocampal cell transduced with two editing variants (CYFIP2 + CYFIP2 K; CYFIP2 + CYFIP2 E) (Tukey’s multiple comparisons test: *p<0.05 ** p<0.01 ***p<0.001 ****p<0.0001).

Article Snippet: pUC19 expressing vector containing the coding sequence of human CYFIP2 (HG16749-U) were acquired from Sino biological Inc. and amplified with primers containing AgeI and SalI restriction sites, used to insert the amplicon into pRRLSIN.cPPT.PGK-GFP.WPRE Lentiviral vector (#12252 Addgene).

Techniques: Derivative Assay, Transduction, shRNA, Confocal Microscopy, Marker, Knockdown