Table 1.

Key resources table

Reagent or resourceSourceIdentifier
Antibodies 
Mouse anti-acetylated tubulin Sigma-Aldrich Cat# T7451, RRID:AB_609894 
Rabbit anti-CP110 Proteintech Cat# 12780-1-AP, RRID:AB_10638480 
Chicken anti-GFP Aves Lab Cat# GFP-100, RRID:AB_2307313 
Rabbit anti-CPAP Proteintech Cat# 11517-1-AP, RRID:AB_2244605 
Rabbit anti-CPAP Kohlmaier et al. (2009)   
Mouse anti-GFP Sigma-Aldrich Cat# G1546 
Rabbit anti-RFP Rockland Immunochemicals Cat# 00–401-379 
Goat anti-mouse Alexa 594 Thermo Fisher Scientific Cat# A-11032, RRID:AB_2534091 
Goat anti-rabbit Atto 647N Sigma-Aldrich Cat# 40839, RRID:AB_1137669 
Goat anti-chicken dylight 488 Thermo Fisher Scientific Cat# SA5–10070, RRID:AB_2556650 
Mouse anti-Ku80 (clone 7) BD Biosciences Cat# 611360, RRID:AB_398882 
Goat anti-mouse IgG2a IRDye 680LT LI-COR Biosciences Cat#926–68051 
Goat anti-rabbit IgG IRDye 800 CW LI-COR Biosciences Cat#926–3221 
Chemicals, peptides, and recombinant proteins 
Thymidine Sigma-Aldrich Cat # T9250-25G 
RO-3306 Selleckchem Cat# S7747 
S-trityl-L-cysteine (STLC) Sigma-Aldrich Cat# 164739-5G 
Puromycin Invivogen Cat# ant-pr-1 
Blasticidin Invivogen Cat# ant-bl-1 
StrepTactin sepharose high performance GE Healthcare Cat# 28–9355-99 
ChromoTek GFP-Trap magnetic particles M-270 ProteinTech Cat# gtma 
Polyethyleneimine hydroscloride (PEI MAX) Polysciences Cat# 24765-1 
cOmplete, EDTA-free protease inhibitor cocktail Roche Cat# 4693116001 
PhosSTOP Roche/Sigma-Aldrich Cat# 4906845001 
Tubulin protein porcine brain Cytoskeleten Cat# T240-C 
Tubulin protein (rhodamine) porcine brain Cytoskeleten Cat# TL590M-B 
Tubulin protein (HiLyte 488 ) porcine brain Cytoskeleten Cat# TL488M-B 
Tubulin protein (HiLyte 647 ) porcine brain Cytoskeleten Cat# TL670M-B 
Tubulin protein biotin porcin brain Cytoskeleten Cat# T333P-B 
GMPCPP Jena Bioscience Cat# NU-405L 
GTP Sigma-Aldrich Cat# G8877 
Glucose oxidase Sigma-Aldrich Cat# G7141 
Catalase Sigma-Aldrich Cat# C9322 
DTT Sigma-Aldrich Cat# R0861 
Κ-casein Sigma-Aldrich Cat# C0406 
Neutravidin Invitrogen Cat# A-2666 
GFP-EB3 full length Montenegro Gouveia et al. (2010)  N/A 
DARPin (TM-3)2 Ahmad et al. (2016), Pecqueur et al. (2012)  N/A 
HisTrap FF 5 ml GE Healthcare Cat #17531901 
Superdex 75 10/30 GL Cytiva Cat #17517401 
HiLoad 16/60 superdex 75 pg column GE Healthcare Cat #28989333 
Superdex 200 increase 10/300 GL GE Healthcare Cat #28990944 
Cell lines 
Human embryonic kidney 239T (HEK293T) cells ATCC Cat #CRL-321 
Flp-In T-Rex U2OS cells Sharma et al. (2016)   
Flp-In U2OS GFP-CPAP-FLWT cells This study N/A 
Flp-In U2OS GFP-CPAP-FLMUT cells This study N/A 
Flp-In U2OS GFP-CPAP-FLWT cells-CPAP KO This study N/A 
Flp-In U2OS GFP-CPAP-FLMUT cells-CPAP KO This study N/A 
Oligonucleotides 
sgRNA-targeting CPAP intron 2 #1 This study 5′-GTG​GCT​GAT​TTA​GTT​ACC​TAG-3′ 
sgRNA-targeting CPAP intron 3 #2 This study 5′-GGT​TCT​TCA​GCT​GAA​CAG​AG-3′ 
CPAP KO-sequencing primer F This study 5′-AAA​AAG​GGA​CCA​CAG​GTA​GCG-3′ 
CPAP KO-sequencing primer R This study 5′-GCT​CCA​GGT​CAC​ATT​TCC​AGT-3′ 
Flp-In siCPA-presistant left part F This study 5′-ACT​TAA​GCT​TGG​TAC​CGA​GCT​CGG​CCA​CCA​TGG​TGA​GCA​AG-3′ 
Flp-In siCPA-presistant left part R This study 5′-CTC​TTC​GAT​CCT​AGC​TAA​CTC​TTT​TGC​TTT​CTG​TTG​TTC​GAA​GT-3′ 
Flp-In siCPA-presistant right part F This study 5′-AAA​AGA​GTT​AGC​TAG​GAT​CGA​AGA​GTT​TAA​AAA​GGA​GGA​GAT​GAG​GA-3′ 
Flp-In siCPA-presistant right part R This study 5′-TTA​AAC​GGG​CCC​TCT​AGA​CTC​GAG​CTC​ACA​GCT​CCG​TGT​CCA​TTA​GC-3′ 
BBNF-mut-CPAP-149–150 This study 5′-AAA​AAC​TTG​AAC​AGG​CCG​CCG​AAG​TAC​AAC​AGA​AGA​AGC​AGG​AAC​AAT​TGA​A-3′, 
BBNR-mut-CPAP-149–150 This study 5′-ATG​GGC​AGG​TAG​CCG​GAT​CAA​GCG​TAT​GCA​GCC​GCC​GCA​T-3′ 
INSF-mut-CPAP-149–-150 This study 5′-ATG​CGG​CGG​CTG​CAT​ACG​CTT​GAT​CCG​GCT​ACC​TGC​CCA​T-3′, 
INSR-mut-CPAP-149–150 This study 5′-TGC​TTC​TTC​TGT​TGT​ACT​TCG​GCG​GCC​TGT​TCA​AGT​TTT​T-3′ 
PX459v2-P2A-BSD F This study 5′-GGC​CAG​GCA​AAA​AAG​AAA​AAG​GAA​TTC​GGC​AGT​GGA​GCT​ACT​AAC​TTC​AGC​CTG​CTG​AAG-3′ 
PX459v2-P2A-BSD R This study 5′-GCT​GAT​CAG​CGA​GCT​CTA​GTT​AGA​ATT​CTC​AGC​CCT​CCC​ACA​CAT​AAC​C-3′ 
2bsai-BSD F This study 5′-CAC​CTG​AGA​CCG​AGG​TCT​CT-3′ 
2bsai-BSD R This study 5′-AAA​CAG​AGA​CCT​CGG​TCT​CA-3′ 
Recombinant DNA 
pcDNA5 FRT TO Thermo Fisher Scientific Cat# V652020 
pcDNA5-FRT-TO-GFP-CPAP-FLWT (plasmid) This study N/A 
pcDNA5-FRT-TO-GFP-CPAP-FLMUT (plasmid) This study N/A 
pOG44 Flp-recombinase expression vector Thermo Fisher Scientific Cat#: V600520 
PX459-sgRNA–targeting CPAP intron 2 (plasmid) This study N/A 
PX459-sgRNA targeting CPAP intron 3 (plasmid) This study N/A 
pSpCas9(BB)–2A-Puro (PX459) V2.0 Ran et al. (2013)  Addgene, RRID: Addgene_62988 
pSpCas9(BB)–2A-BSD (PX459-BSD) This study N/A 
pTT5-SII-GFP-CP110 This study N/A 
pTT5-CEP97-GFP-SII This study N/A 
pTT5-CEP97(1-650)^CP110(581-991)-GFP-SII This study N/A 
pTT5-CPAP(1-607)WT-mCherry-SII This study N/A 
pTT5-CPAP(1-607)L149A/K150A-mCherry-SII This study N/A 
PSTCm9 Olieric et al. (2010)  N/A 
Software and algorithms 
FIJI (ImageJ) National Institutes of Health https://fiji.sc/; https://imagej.net/ij/index.html; RRID: SCR_003070 
GraphPad prism 9.0.0 (121) GraphPad Software RRID: SCR_000306 
Adobe Illustrator 2023 Adobe Systems RRID: SCR_010279 
Huygens professional 21.04 Scientific Volume Imaging B.V. https://svi.nl 
Metamorph Molecular Devices https://www.moleculardevices.com/products/cellular-imaging-systems/high-content-analysis/metamorph-microscopy 
KymoResliceWide plugin Eugene Katrukha https://github.com/ekatrukha/KymoResliceWide 
MATLAB code Eugene Katrukha https://gist.github.com/ekatrukha/8a8f336d44b7bef6523fc1acc3c71a19 
UCSF chimera Pettersen et al. (2004)  https://www.cgl.ucsf.edu/chimera/ 
Situs 3 Kovacs et al. (2018)   
DAMAVER Volkov and Svergun (2003)  https://www.embl-hamburg.de/biosaxs/damaver.html 
DAMMIF Franke and Svergun (2009)  https://www.embl-hamburg.de/biosaxs/dammif.html 
Scatter3 Forster et al. (2010)   
PyMol Schrödinger https://pymol.org/ 
Reagent or resourceSourceIdentifier
Antibodies 
Mouse anti-acetylated tubulin Sigma-Aldrich Cat# T7451, RRID:AB_609894 
Rabbit anti-CP110 Proteintech Cat# 12780-1-AP, RRID:AB_10638480 
Chicken anti-GFP Aves Lab Cat# GFP-100, RRID:AB_2307313 
Rabbit anti-CPAP Proteintech Cat# 11517-1-AP, RRID:AB_2244605 
Rabbit anti-CPAP Kohlmaier et al. (2009)   
Mouse anti-GFP Sigma-Aldrich Cat# G1546 
Rabbit anti-RFP Rockland Immunochemicals Cat# 00–401-379 
Goat anti-mouse Alexa 594 Thermo Fisher Scientific Cat# A-11032, RRID:AB_2534091 
Goat anti-rabbit Atto 647N Sigma-Aldrich Cat# 40839, RRID:AB_1137669 
Goat anti-chicken dylight 488 Thermo Fisher Scientific Cat# SA5–10070, RRID:AB_2556650 
Mouse anti-Ku80 (clone 7) BD Biosciences Cat# 611360, RRID:AB_398882 
Goat anti-mouse IgG2a IRDye 680LT LI-COR Biosciences Cat#926–68051 
Goat anti-rabbit IgG IRDye 800 CW LI-COR Biosciences Cat#926–3221 
Chemicals, peptides, and recombinant proteins 
Thymidine Sigma-Aldrich Cat # T9250-25G 
RO-3306 Selleckchem Cat# S7747 
S-trityl-L-cysteine (STLC) Sigma-Aldrich Cat# 164739-5G 
Puromycin Invivogen Cat# ant-pr-1 
Blasticidin Invivogen Cat# ant-bl-1 
StrepTactin sepharose high performance GE Healthcare Cat# 28–9355-99 
ChromoTek GFP-Trap magnetic particles M-270 ProteinTech Cat# gtma 
Polyethyleneimine hydroscloride (PEI MAX) Polysciences Cat# 24765-1 
cOmplete, EDTA-free protease inhibitor cocktail Roche Cat# 4693116001 
PhosSTOP Roche/Sigma-Aldrich Cat# 4906845001 
Tubulin protein porcine brain Cytoskeleten Cat# T240-C 
Tubulin protein (rhodamine) porcine brain Cytoskeleten Cat# TL590M-B 
Tubulin protein (HiLyte 488 ) porcine brain Cytoskeleten Cat# TL488M-B 
Tubulin protein (HiLyte 647 ) porcine brain Cytoskeleten Cat# TL670M-B 
Tubulin protein biotin porcin brain Cytoskeleten Cat# T333P-B 
GMPCPP Jena Bioscience Cat# NU-405L 
GTP Sigma-Aldrich Cat# G8877 
Glucose oxidase Sigma-Aldrich Cat# G7141 
Catalase Sigma-Aldrich Cat# C9322 
DTT Sigma-Aldrich Cat# R0861 
Κ-casein Sigma-Aldrich Cat# C0406 
Neutravidin Invitrogen Cat# A-2666 
GFP-EB3 full length Montenegro Gouveia et al. (2010)  N/A 
DARPin (TM-3)2 Ahmad et al. (2016), Pecqueur et al. (2012)  N/A 
HisTrap FF 5 ml GE Healthcare Cat #17531901 
Superdex 75 10/30 GL Cytiva Cat #17517401 
HiLoad 16/60 superdex 75 pg column GE Healthcare Cat #28989333 
Superdex 200 increase 10/300 GL GE Healthcare Cat #28990944 
Cell lines 
Human embryonic kidney 239T (HEK293T) cells ATCC Cat #CRL-321 
Flp-In T-Rex U2OS cells Sharma et al. (2016)   
Flp-In U2OS GFP-CPAP-FLWT cells This study N/A 
Flp-In U2OS GFP-CPAP-FLMUT cells This study N/A 
Flp-In U2OS GFP-CPAP-FLWT cells-CPAP KO This study N/A 
Flp-In U2OS GFP-CPAP-FLMUT cells-CPAP KO This study N/A 
Oligonucleotides 
sgRNA-targeting CPAP intron 2 #1 This study 5′-GTG​GCT​GAT​TTA​GTT​ACC​TAG-3′ 
sgRNA-targeting CPAP intron 3 #2 This study 5′-GGT​TCT​TCA​GCT​GAA​CAG​AG-3′ 
CPAP KO-sequencing primer F This study 5′-AAA​AAG​GGA​CCA​CAG​GTA​GCG-3′ 
CPAP KO-sequencing primer R This study 5′-GCT​CCA​GGT​CAC​ATT​TCC​AGT-3′ 
Flp-In siCPA-presistant left part F This study 5′-ACT​TAA​GCT​TGG​TAC​CGA​GCT​CGG​CCA​CCA​TGG​TGA​GCA​AG-3′ 
Flp-In siCPA-presistant left part R This study 5′-CTC​TTC​GAT​CCT​AGC​TAA​CTC​TTT​TGC​TTT​CTG​TTG​TTC​GAA​GT-3′ 
Flp-In siCPA-presistant right part F This study 5′-AAA​AGA​GTT​AGC​TAG​GAT​CGA​AGA​GTT​TAA​AAA​GGA​GGA​GAT​GAG​GA-3′ 
Flp-In siCPA-presistant right part R This study 5′-TTA​AAC​GGG​CCC​TCT​AGA​CTC​GAG​CTC​ACA​GCT​CCG​TGT​CCA​TTA​GC-3′ 
BBNF-mut-CPAP-149–150 This study 5′-AAA​AAC​TTG​AAC​AGG​CCG​CCG​AAG​TAC​AAC​AGA​AGA​AGC​AGG​AAC​AAT​TGA​A-3′, 
BBNR-mut-CPAP-149–150 This study 5′-ATG​GGC​AGG​TAG​CCG​GAT​CAA​GCG​TAT​GCA​GCC​GCC​GCA​T-3′ 
INSF-mut-CPAP-149–-150 This study 5′-ATG​CGG​CGG​CTG​CAT​ACG​CTT​GAT​CCG​GCT​ACC​TGC​CCA​T-3′, 
INSR-mut-CPAP-149–150 This study 5′-TGC​TTC​TTC​TGT​TGT​ACT​TCG​GCG​GCC​TGT​TCA​AGT​TTT​T-3′ 
PX459v2-P2A-BSD F This study 5′-GGC​CAG​GCA​AAA​AAG​AAA​AAG​GAA​TTC​GGC​AGT​GGA​GCT​ACT​AAC​TTC​AGC​CTG​CTG​AAG-3′ 
PX459v2-P2A-BSD R This study 5′-GCT​GAT​CAG​CGA​GCT​CTA​GTT​AGA​ATT​CTC​AGC​CCT​CCC​ACA​CAT​AAC​C-3′ 
2bsai-BSD F This study 5′-CAC​CTG​AGA​CCG​AGG​TCT​CT-3′ 
2bsai-BSD R This study 5′-AAA​CAG​AGA​CCT​CGG​TCT​CA-3′ 
Recombinant DNA 
pcDNA5 FRT TO Thermo Fisher Scientific Cat# V652020 
pcDNA5-FRT-TO-GFP-CPAP-FLWT (plasmid) This study N/A 
pcDNA5-FRT-TO-GFP-CPAP-FLMUT (plasmid) This study N/A 
pOG44 Flp-recombinase expression vector Thermo Fisher Scientific Cat#: V600520 
PX459-sgRNA–targeting CPAP intron 2 (plasmid) This study N/A 
PX459-sgRNA targeting CPAP intron 3 (plasmid) This study N/A 
pSpCas9(BB)–2A-Puro (PX459) V2.0 Ran et al. (2013)  Addgene, RRID: Addgene_62988 
pSpCas9(BB)–2A-BSD (PX459-BSD) This study N/A 
pTT5-SII-GFP-CP110 This study N/A 
pTT5-CEP97-GFP-SII This study N/A 
pTT5-CEP97(1-650)^CP110(581-991)-GFP-SII This study N/A 
pTT5-CPAP(1-607)WT-mCherry-SII This study N/A 
pTT5-CPAP(1-607)L149A/K150A-mCherry-SII This study N/A 
PSTCm9 Olieric et al. (2010)  N/A 
Software and algorithms 
FIJI (ImageJ) National Institutes of Health https://fiji.sc/; https://imagej.net/ij/index.html; RRID: SCR_003070 
GraphPad prism 9.0.0 (121) GraphPad Software RRID: SCR_000306 
Adobe Illustrator 2023 Adobe Systems RRID: SCR_010279 
Huygens professional 21.04 Scientific Volume Imaging B.V. https://svi.nl 
Metamorph Molecular Devices https://www.moleculardevices.com/products/cellular-imaging-systems/high-content-analysis/metamorph-microscopy 
KymoResliceWide plugin Eugene Katrukha https://github.com/ekatrukha/KymoResliceWide 
MATLAB code Eugene Katrukha https://gist.github.com/ekatrukha/8a8f336d44b7bef6523fc1acc3c71a19 
UCSF chimera Pettersen et al. (2004)  https://www.cgl.ucsf.edu/chimera/ 
Situs 3 Kovacs et al. (2018)   
DAMAVER Volkov and Svergun (2003)  https://www.embl-hamburg.de/biosaxs/damaver.html 
DAMMIF Franke and Svergun (2009)  https://www.embl-hamburg.de/biosaxs/dammif.html 
Scatter3 Forster et al. (2010)   
PyMol Schrödinger https://pymol.org/ 
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