Table 2 Identification of single-cell clones bearing NanoLuc integrations in correct reading frame.

From: A generic strategy for CRISPR-Cas9-mediated gene tagging

  

PCR positive

In-frame integration

Gene

gRNA sequence

5′ junction

Both junctions

5′ junction

Both junctions

IFIT1

5′-CCAATTTGTAGACGAACCCA-3′

7/84

4/7

2/4

1/2

DACT1

5′-ACAAGCGAACTGACTACCGG-3′

5/96

3/5

3/3

0/3

EGR1

5′-GTACGTGGTGGCCACCGACG-3′

2/96

1/2

1/1

1/1

  1. gRNA, guide RNA.
  2. HAP1 cells were transfected with expression plasmids for Cas9, the generic NanoLuc donor specified in Supplementary Fig. 3 and one gRNA for each gene under consideration (IFIT1, DACT1, EGR1). Single-cell clones were isolated by limiting dilution and analysed by PCR and Sanger sequencing.
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