Fig. 7: SPINDOC facilitates PARP1-mediated PARylation and targets transcription in vivo.

a Schematic of the establishment of SPINDOC KO mice. Mus musculus SPINDOC sgRNA is shown and Pam site is highlighted in red. Founders were backcrossed to the F3 generation before being intercrossed to generate KO mice. The model was created with BioRender.com. b Three independent founders were identified, harboring different out-of-frame indels (red) around the sgRNA sequence (blue). These founders were termed #18, #19, and #21. A PCR genotyping, using primers that straddled the edited regions, was developed. An example of the genotyping of a full litter from each of the three independent lines is shown. c, d SPINDOC KO from line #18 (c) and line #19 (d), and their corresponding WT littermates, were treated with 6-Gy IR and then allowed to recover for 3 h before their thymi were isolated. The thymi were lysed and subjected to Western blot analysis, using the indicated antibodies. The three-digit numbers above each lane refer to the identity number of the mouse from which the thymus was isolated. e, f Mice were treated as in c, d and their thymic tissue was sonicated before total RNA extraction and qRT-PCR analysis. Tissues were taken from sacrificed lines #18 and #19. Graphs represent mean ± SD, n = 3 biologically independent mice. One-tailed Student t-tests were performed. e Line #18, *P = 0.0422, **P = 0.0041; line #19, KO-NT vs WT-NT, P = 0.0109, KO-IR vs WT-NT, P = 0.0127. f Line #18, KO-NT vs WT-NT, P = 0.0024, WT-IR vs WT-NT, P = 0.0161; line #19, KO-NT vs WT-NT, P = 0.0017, KO-IR vs WT-IR, P = 0.0035, WT-IR vs WT-NT, P = 0.0054, KO-IR vs WT-NT, P = 0.0066. ***P < 0.001, ****P < 0.0001. g SPINDOC WT (n = 20) and KO (n = 22) mice were exposed to 6 Gy of IR and maintained for up to 35 days. Statistical significance of survival rates between genotypes was determined using the Kaplan–Meier method and P value was calculated by a log-rank test.