Lentivirus production and stable expression in Ba/F3 cells

Takahiro Yoshizawa; Ken Uchibori; Mitsugu Araki; Shigeyuki Matsumoto; Biao Ma; Ryo Kanada; Yosuke Seto; Tomoko Oh-hara; Sumie Koike; Ryo Ariyasu; Satoru Kitazono; Hironori Ninomiya; Kengo Takeuchi; Noriko Yanagitani; Satoshi Takagi; Kazuma Kishi; Naoya Fujita; Yasushi Okuno; Makoto Nishio; Ryohei Katayama

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Lentivirus production and stable expression in Ba/F3 cells

TY Takahiro Yoshizawa

KU Ken Uchibori

MA Mitsugu Araki

SM Shigeyuki Matsumoto

BM Biao Ma

RK Ryo Kanada

YS Yosuke Seto

TO Tomoko Oh-hara

SK Sumie Koike

RA Ryo Ariyasu

SK Satoru Kitazono

HN Hironori Ninomiya

KT Kengo Takeuchi

NY Noriko Yanagitani

ST Satoshi Takagi

KK Kazuma Kishi

NF Naoya Fujita

YO Yasushi Okuno

MN Makoto Nishio

RK Ryohei Katayama

This method is extracted from research article: NPJ Precis Oncol, Apr 2021

Microsecond-timescale MD simulation of EGFR minor mutation predicts the structural flexibility of EGFR kinase core that reflects EGFR inhibitor sensitivity

DOI: 10.1038/s41698-021-00170-7

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The EGFR-L747P, del19, L858R, del19/T790M, or L858R/T790M mutant was created by Quikchange mutagenesis using pENTR-EGFR, and lentiviral plasmid pLenti6.3-EGFR-L747P or other mutants were cloned by LR clonase II using pENTR-EGFR (L747P or other mutants) and pLenti6.3-DEST (Invitrogen). Lentivirus was prepared by transfecting pLenti6.3-EGFR-L747P or other mutants with a helper plasmid (ViraPower) in 293FT cells. Ba/F3 cells were infected using lentivirus-containing medium supplemented with polybrene (8 µg/mL), and after an incubation of 24 h, the infected cells were selected using 7 µM blasticidin (Invitrogen) for one week. After the selection, cells were cultured in a culture medium without IL-3.

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