| Chemicals, peptides, and recombinant proteins |
|
| DMEM/F-12, supplemented with GlutaMAX™
|
Life Technologies |
Cat#: 10565042 |
| tetracycline-free fetal bovine serum |
Takara Bio |
Cat#: 631106 |
| TrypLE™ Express Enzyme (1X) |
Gibco Life Technologies |
Cat#: 12604039 |
| Hygromycin B (50 mg/mL) |
Gibco Life Technologies |
Cat# 10687010 |
| Blasticidin S (10 mg/mL) |
Gibco Life Technologies |
Cat# A1113903 |
| Doxycycline (1 μg/mL) |
Millipore Sigma |
Cat# D9891-10G |
| TransIT-LT1 (lentivirus production) |
Mirus Bio LCC |
Cat# MIR 2306 |
| Polybrene (10 mg/mL) |
Millipore Corporation |
Cat# TR-1003-G |
| Puromycin dihydrochloride (10 mg/mL) |
Gibco Life Technologies |
Cat# A1113803 |
| Veratridine (CAS: 71-62-5) |
Sigma-Aldrich |
Cat# v5754-25 |
| Ouabain (CAS: 11018-89-6) |
Sigma-Aldrich |
Cat# PHR1945-300MG |
| Lipofectamine 2000 (cell line generation) |
Life Technologies |
Cat# 11668019 |
| Proteinase K (>600 mAU/mL) (gDNA extraction) |
QIAGEN |
Cat# 19133 |
| RNase A (100 mg/mL) (gDNA extraction) |
QIAGEN |
Cat# 19101 |
| DMSO (PCR) |
Sigma Aldrich |
Cat# D9170-5VL |
| Electroporation Buffer (electroporation) |
Maxcyte |
Cat#: EPB-1 |
|
| Critical commercial assays |
|
| Cell Titer Glo |
Promega |
Cat# G7571 |
| Titanium Taq DNA Polymerase |
Clontech Takara |
Cat# 639208 |
| 10X Titanium Taq buffer |
Clontech Takara |
Cat# 639242 |
| PureLink HiPure Plasmid DNA Purification Kit (lentivirus production) |
Invitrogen |
Cat# K2100-04 |
| DNeasy Blood and Tissue Kit (gDNA extraction) |
QIAGEN |
Cat# 69504 |
| dsDNA HS Assay |
Life Technologies |
Cat# Q33230
|
| NEBNext High Fidelity 2X PCR Master Mix |
New England Biolabs |
Cat# M0541 |
| dNTPs |
Clontech Takara |
Cat# 4030 |
| SPRI beads |
Beckman Coulter |
Cat# B23318
|
|
| Deposited data |
|
| Sequencing data from gRNA validation |
This paper |
https://doi.org/10.5061/dryad.zkh1893fc
|
|
| Experimental models: Cell lines |
|
| Stable Nav1.2 WT Cell Line (background FlpIn TREx HEK293 cells) |
This paper (hSCN2A: NM_021007) |
N/A |
| HEK293T |
ATCC |
Cat# CRL-3216 |
|
| Oligonucleotides |
|
| See Tables S4 and S5
|
N/A |
N/A |
|
| Recombinant DNA |
|
| pRDA_256 (for library and variant guides) |
Hanna et al.25 (The Broad Institute) |
Addgene #158581 |
| psPAX2 (for lentivirus production) |
Hanna et al.25 (The Broad Institute) |
Addgene #12260 |
| pCMV_VSVG (for lentivirus production) |
Hanna et al.25 (The Broad Institute) |
Addgene #8454 |
| pIR-CMV-SCN2A-Variant-1-IRES-mScarlet |
DeKeyser et al.36 (SCN2A: NM_021007) |
Addgene #162279 |
|
| Software and algorithms |
|
| Prism7 |
GraphPad Software |
https://www.graphpad.com/
|
| CRISPResso2 (version 2.1.3) |
Clement et al.37
|
https://github.com/pinellolab/CRISPResso2
|
| Biorender |
Biorender |
https://app.biorender.com/
|
| DataControl384 (Syncropatch) |
Nanion |
https://www.nanion.de/products/syncropatch-384/software/
|
| SnapGene software |
SnapGene by Dotmatics |
www.snapgene.com
|
| Our Code |
GitHub |
https://doi.org/10.5281/zenodo.7901798
|
