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. Author manuscript; available in PMC: 2020 Aug 26.
Published in final edited form as: Cell Rep. 2020 Jul 14;32(2):107878. doi: 10.1016/j.celrep.2020.107878

Table 1.

Editing of Adenosines within Mecp2 RNA Identified by Sanger Sequencing Analysis

graphic file with name nihms-1612482-t0002.jpg
Target
DG Replicate Injection Condition E102G E102E T105A T105T R106Qa K107R
1 6xU6-Mecp2 targeting guide-hSynI editase ND 44.5% ND ND 39.4% ND
2 6xU6-Mecp2 targeting guide-hSynI editase 10.2% 41.3% 15.3% 9.4% 57.9% 9.6%
3 6xU6-Mecp2 targeting guide-hSynI editase 7.8% 39.2% 8.3% ND 49.7% ND
1 1xU6-non-targeting guide-hSynI editase ND ND ND ND ND ND
2 1xU6-non-targeting guide-hSynI editase ND ND ND ND ND ND
3 1xU6-non-targeting guide-hSynI editase ND ND ND ND ND ND
CA1 Replicate Injection Condition E102G E102E T105A T105T R106Qa K107R
1 6xU6-Mecp2 targeting guide-hSynI editase 5.8% 64.7% ND ND 63.9% 64.7%
2 6xU6-Mecp2 targeting guide-hSynI editase ND 24.4% 6% ND 39.6% ND
3 6xU6-Mecp2 targeting guide-hSynI editase ND 18.3% ND ND 52.4% ND
1 1xU6-non-targeting guide-hSynI editase ND ND ND ND ND ND
2 1xU6-non-targeting guide-hSynI editase ND ND ND ND ND ND
3 1xU6-non-targeting guide-hSynI editase ND ND ND ND ND ND
CA3 Replicate Injection Condition E102G E102E T105A T105T R106Qa K107R
1 6xU6-Mecp2 targeting guide-hSynI editase 5.9% 38% 16.3% 8.4% 41% 6.2%
2 6xU6-Mecp2 targeting guide-hSynI editase 9.3% 39.6% 18.9% 12.1% 54.9% 12.9%
3 6xU6-Mecp2 targeting guide-hSynI editase 7.5% 26.8% 12.7% 9.8% 49.7% ND%
1 1xU6-non-targeting guide-hSynI editase ND ND ND ND ND ND
2 1xU6-non-targeting guide-hSynI editase ND ND ND ND ND ND
3 1xU6-non-targeting guide-hSynI editase ND ND ND ND ND ND

Top: Mecp2 mRNA and the deduced primary amino acid sequence (top row) relative to the guide RNA (bottom row). x, mismatched adenosines. Bottom: rates of editing at the adenosines located within the guide region for each biological replicate. The detection limit for this assay was determined previously to be 5% editing (Sinnamon et al., 2017). Any sites that had 5% or less editing are listed as having no detectable editing (ND). There was no detectable editing in Mecp2 RNA outside of the guide region.

a

The target adenosine, Mecp2317 in the RNA strand (MeCP2R106Q).