(A) Simple illustration of how Pou4f3-/- strain was generated. (B) One example gel image of tail-DNA PCR used to distinguish WT (Pou4f3+/+), heterozygous (Pou4f3+/-), and homozygous (Pou4f3-/-) …
The original agarose gel image in Figure 1B (Pou4f3+/+, Pou4f3+/-, and Pou4f3-/-).
File containing Figure 1B and the original agarose gel analysis with highlighted bands and sample labels.
(A) Illustration of how Pou4f3-/- mice (Founder 0, F0) were rapidly generated using CRISPR-stop. (B–, C) Sanger sequencing chromatograms of WT (B) and F0 Pou4f3-/- (C) samples. The base ‘C’ (black …
(A–C’’) Dual staining of POU4F3 and MYO7A in middle-turn regions of cochleae from WT mice (A–A’’) at P1 and Pou4f3-/- mice at P0 (B–B’’) and P1 (C–C’’). The Pou4f3-/- mice were from a …
File containing raw counts of the cochlear hair cells reported in the graph of Figure 1—figure supplement 2D.
(A) Simple cartoon depicting how Gfi1 was inactivated in one-cell-stage zygotes derived from male Gfi1HA-Cre/+ mice by using the CRISPR-stop approach. (B, C) Using sgRNA-6 as an example, Sanger …
(A–C) Simple cartoon illustrating construction of Gfi1HA-Cre/+ strain. In the WT Gfi1 allele (A), a DNA fragment containing 3×HA-P2A-Cre (B) was inserted immediately before the TGA stop codon (red …
The original agarose gel image in Figure 2—figure supplement 1D (Gfi1+/+, Gfi1HA-Cre/+, and Gfi1HA-Cre/ HA-Cre).
File containing Figure 2—figure supplement 1D and the original agarose gel analysis with highlighted bands and sample labels.
(A) One-step generation of Pou4f3 mutants by using CRISPR-stop with zygotes derived from Gfi1HA-Cre/+ mice. (B, C) Sanger sequencing chromatograms of tail-DNA samples from control Gfi1HA-Cre/+ (B) …
(A) The DNA fragment between sgRNA-4 and sgRNA-9 within the Gfi1 locus was deleted using the CRISPR/Cas9 method. (B) One example gel image of tail-DNA PCR used to distinguish Gfi1+/+, Gfi1+/-, and Gf…
The original agarose gel image in Figure 2—figure supplement 3B (Gfi1+/+, Gfi1+/-, and Gfi1-/-).
File containing Figure 2—figure supplement 3B and the original agarose gel analysis with highlighted bands and sample labels.
(A) Simple cartoon showing how CRISPR-stop was used to induce mosaic inactivation of RBM24 in one-cell-stage mouse zygotes harboring Gfi1HA-Cre/+. Notably, the injection was in the late one-cell …
(A) Simple illustration of how mosaic Rbm24 mutant mice were generated. (B–C’’) Dual staining of RBM24 and GATA3 in cochlear tissues from control Rbm24+/+ (B–B’’) and Rbm24-/- (C–C’’) mice at P1. (D)…
(A) Visualization of ATAC-seq (top row, orange) of P1 cochlear HCs (from our previous study, GSE181311) and POU4F3 Cut&Run assay (bottom row, blue) of E17.5 cochlear HCs (from another study, …
(A) Generation of Rosa26loxp-stop-loxp-Rbm24*3×HA/+ (Rosa26Rbm24/+) mouse strain. The C-terminus of ectopic RBM24 was fused with three HA tags. (B) One example gel image of tail-DNA PCR used to …
The original agarose gel image in Figure 5B (Rosa26+/+ and Rosa26Rbm24/+).
File containing Figure 5B and the original agarose gel analysis with highlighted bands and sample labels.
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Strain, strain background (Mus musculus) | Atoh1Cre/+ | Yang et al., 2010b | Lin Gan lab (Augusta University) | A knock-in mouse strain |
Strain, strain background (M. musculus) | Rosa26-loxp-stop-loxp-tdTomato (Ai9) | Stock# 007905 | The Jackson Laboratory | A knock-in mouse strain |
Strain, strain background (M. musculus) | Ikzf2V5/+ | Stock# 038463 | The Jackson Laboratory | A knock-in mouse strain |
Strain, strain background (Mus musculus) | Pou4f3 mutant | This paper | Available upon request from Liu Lab | Constructed by CRISPR-stop |
Strain, strain background (M. musculus) | Gfi1 mutant | This paper | Available upon request from Liu Lab | Constructed by CRISPR-stop |
Strain, strain background (M. musculus) | Rbm24 mutant | This paper | Available upon request from Liu Lab | Constructed by CRISPR-stop |
Strain, strain background (M. musculus) | Pou4f3 null | This paper | Available upon request from Liu Lab | Constructed by CRISPR /Cas9 |
Strain, strain background (M. musculus) | Gfi1 null | This paper | Available upon request from Liu Lab | Constructed by CRISPR /Cas9 |
Strain, strain background (M. musculus) | Gfi13xHA-P2A-Cre | This paper | Available upon request from Liu Lab | A knock-in mouse strain |
Strain, strain background (M. musculus) | Eh1-EGFP+ | This paper | Available upon request from Liu Lab | A transgenic mouse line |
Strain, strain background (M. musculus) | Eh2-EGFP+ | This paper | Available upon request from Liu Lab | A transgenic mouse line |
Strain, strain background (M. musculus) | Eh3-EGFP+ | This paper | Available upon request from Liu Lab | A transgenic mouse line |
Strain, strain background (M. musculus) | Rosa26CAG-lsl-Rbm24*3xHA/+ | This paper | Available upon request from Liu Lab | A knock-in mouse strain for ectopic Rbm24 expression |
Antibody | Anti-RBM24 (rabbit polyclonal) | Proteintech | 18178-1-AP | IF (1:500) |
Antibody | Anti-POU4F3 (mouse monoclonal) | Santa Cruz | sc-81980 | IF (1:500) |
Antibody | Anti-POU4F3 (rabbit polyclonal) | Novus Biologicals | NBP1-88349 | IF (1:500) |
Antibody | Anti-HA (rat monoclonal) | Roche | 11867423001 | IF (1:500) |
Antibody | Anti-MYO7A (rabbit polyclonal) | Proteus Biosciences | 25-6790 | IF (1:500) |
Antibody | Anti-MYO7A (mouse monoclonal) | DSHB | MYO7A 138-1 | IF (1:500) |
Antibody | Anti-CTIP2 (rat monoclonal) | Abcam | ab18465 | IF (1:500) |
Antibody | Anti-INSM1 (guinea pig) | Jia et al., 2015 | A kind gift from Dr. Carmen Birchmeier | IF (1:6000) |
Antibody | Anti-GFP (chicken polyclonal) | Abcam | ab13970 | IF (1:500) |
Antibody | Anti-GATA3 (goat polyclonal) | R&D Systems | AF2605 | IF (1:500) |
Antibody | Anti-V5 (mouse monoclonal) | Bio-Rad | MCA1360 | IF (1:500) |
List of sgRNA sequences.
DNA sequences of ssDNA donor and Rbm24 enhancers.
DNA sequences of genotyping primers.