ENCAB490KGN

Homo sapiens

any cell type or tissue

partially characterized

Method: knockdown or knockout

compliant

Caption: Western blot following CRISPR against SEC62 in K562 whole cell lysate using SEC62 specific antibody. Lane 1 is a ladder, lane 2 is K562 non-targeting control knockdown, lane 3 and 4 are two different CRISPR against SEC62. SEC62 protein appears as the green arrow, Beta-actin serves as a control and appears in red arrow.
Reviewer comment: Checked with Eric Mendenhall and he agreed with the lab’s explanation: Based on the explanation we got from the lab, these are all CRISPRi based knockouts (unlike the previous RNAi based knockouts) where the lab introduces a stop codon, thus preventing protein synthesis altogether. And as a result, they are able to detect only the protein knockdown rates, but the RNA levels still might not be changing and thus they report 0% k/d using qRT-PCR. This is unlike the RNAi based silencing methods.
Submitted by: Xintao Wei
Lab: Brenton Graveley, UConn
Grant: U41HG009889
Download: SEC62-K562-CRISPR-A303-981A.png
Documents: encode:TF_Antibody_Characterization_ENCODE3_May2016.pdf