HEK293T-CAF40-null

ACC 872

Cell line:

HEK293T-CAF40-null

DSMZ no.:

ACC 872

Species:

human (Homo sapiens)

Cell type:

embryonal kidney

Origin:

this cell line is part of a set of cell lines models carrying mutations in different genes affecting mRNA processing and translation efficiency (ACC 872 - ACC 876). Single clone derivative of 293T (DSMZ ACC 635) which was modified by CRISPR/CAS9 editing. In this cell line the ORF of the CNOT9 (CAF40) gene was disrupted by random frameshift mutation in exon 2. The CCR4-NOT transcription complex subunit 9 (CNOT9, also known as CAF40) is a multiprotein complex regulating gene expression via shortening of poly(A) tails of mRNA or by deadenylation.

Reference(s):

18207, 18209

Biosafety level:

1, GMO-S1

Risk assessment:

genetically modified organism, according to German and European legislation allocated into group S1 (see also parental 293T, DSMZ: ACC 635)

Permissions and restrictions:

A, D

Depositor-specific restrictions:

Transfer to third parties needs to be negotiated with the depositor.

DSMZ Cell Culture Data:

Morphology:

oval to fibroblastoid cells growing loosely adherently as monolayer; image; image

Medium:

90% DMEM + 10% h.i. FBS

Subculture:

seed out at about 1-2 x 10⁶ cells/25 cm²; split (semi-)confluent culture 1:2 to 1:3 every second day by tapping the flask (trypsin/EDTA is not necessary)

Incubation:

at 37 °C with 5% CO₂

Doubling time:

ca. 30-40 hours

Harvest:

cell harvest of ca. 20 x 10⁶ cells/80 cm² flask

Storage:

frozen with 70% medium, 20% FBS, 10% DMSO

DSMZ Scientific Data:

Mycoplasma:

negative in PCR assay

Immunology:

to inquire about expression of EpCAM and intermediate filaments, contact ulfert.rand@dsmz.de.

Authenticity:

STR analysis according to the global standard ANSI/ATCC ASN-0002.1-2021 (2021) resulted in an authentic STR profile of the reference STR database.

Molec. Genetics:

disruption of the CNOT9 (CAF40) gene was verified by sequence analysis as described in Ref 18207 (Sgromo et al 2018). For internal control the following primers can be used D-CNOT9-F: AACGCTCCCCTGGTATTCATGAAA and D-CNOT9-R: TTAAAGAATCATCCTCCTAAAGTCTTCAAATC. Absence of the CNOT9 protein was confirmed by Western blot.

Viruses:

PCR: EBV -, HBV -, HCV -, HIV-1 -, HIV-2 -, HTLV-1/2 -, MLV -

Supplied as:

Delivery form		Prices

Frozen culture		450,- €
Growing culture (please inquire for exact delivery time)		900,- €
DNA isolated from cell line (25 µg)		560,- €
DNA isolated from cell line (5 µg)		140,- €

see price list

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