REAGENT or RESOURCE	SOURCE	IDENTIFIER
Bacterial and virus strains
T1 competent cells	Shanghai Weidi Biotechnology Co, Ltd	DL1015S
Chemicals, peptides, and recombinant proteins
Fetal bovine serum (FBS)	Sigma, USA	F2442
Pennicillin/Streptomycin solution	Beyotime, China	C0222
Trypsin-EDTA solution	Sigma, USA	T4049
Leukemia inhibitory factor (LIF) or ESGRO	Millipore, USA	ESG1107
T4 DNA ligase	Takara, Japan	2011A
T4 polynucleotide kinase (PNK) (10 U/ μL)	Takara, Japan	2021S
Bpil	Thermo Scientific, USA	FD1014
Gelatin	Sinopharm Chemical Reagent (SCR), China	9000-70-8
Bacto-yeast extract	Sangon, China	A515245-0500
Bacto-tryptone	Sangon, China	A505247-0500
Agar	Macklin, China	A800730
Lipofectamine 2000	Invitrogen, USA	11668019
DMEM	Invitrogen, USA	C11995500CP
Non-essential amino acid (NEAA) solution (100×)	HyClone, USA	SH30238.01
Dithiothreitol (DTT)	Fisher Biotec, USA	BP172-25
Dimethyl sulfoxide (DMSO)	Amethyst, China	966629
Carbenicillin	YEASEN, China	60202ES08
β-mercaptoethanol	Sigma, USA	M3148
84 disinfection solution (with sodium hypochlorite)	Aitefu, China	92651097
Sodium Chloride (NaCl)	Sangon, China	A501218-0001
PBS (20×)	Sangon, China	B548117-0500
Proteinase K	Abcone, China	P78893
Tris	Beyotime Biotechnology, China	ST761
SDS	Sigma, USA	L4390
Agarose	Abcone, China	A88490
GeneGreen nucleic acid dye	TIANGEN, China	RT210
EDTA	Amethyst, China	976151-500G
EcoRV	NEB, USA	R0195S
HindIII	NEB, USA	R0104S
Critical commercial assays
T7E1 assay	Vazyme, China	EN303-01
Experimental models: Mouse wild-type ES cell lines
D1911 ES cell line (129/DBA hybrid background)	Li Lab, ShanghaiTech	(Lau et al., 2016)
D1906 ES cell line (129/DBA hybrid background)	Li Lab, ShanghaiTech	(Lau et al., 2016)
TC1 ES cell line (129 background)	Leder Lab, Harvard	(Deng et al., 1995)
Oligonucleotides
CACCGAATAGGAATTTGTGACGTCC	GENEWIZ	Zfp445-sgRNA1-top
AAACGGACGTCACAAATTCCTATTC	GENEWIZ	Zfp445-sgRNA1-bottom
CACCGAGCTCAGCGCAATCTTTATC	GENEWIZ	Zfp445-sgRNA2-top
AAACGATAAAGATTGCGCTGAGCT	GENEWIZ	Zfp445-sgRNA2-bottom
AGTGCGTCCTTCGTTACCTG	GENEWIZ	Zfp445-F1
GTGAAGGTAGCTGGGGATAC	GENEWIZ	Zfp445-R1
GAGGGCCTATTTCCCATGATTCCT	GENEWIZ	U6-Forward
GGACCTACAAGCAAGCAGAC	GENEWIZ	Zfp445-F2
GTCACCGAGCTGCAAGAACT	GENEWIZ	Puro-F
GCTCGTAGAAGGGGAGGTTG	GENEWIZ	Puro-R
GGAAACAGCTATGACCATG	GENEWIZ	M13 Reverse Primer
Recombinant DNA
pX330	Addgene, USA	42230
PGL3-U6-sgRNA-PGK-puromycin	Youbio, China	VT8203
pBluescript	GenBank	X52331.1
Other
AxyPrep DNA Gel Extraction Kit	Axygen of Corning	35717KE1
AxyPrep Endo-Free Plasmid Midiprep Kit	Axygen of Corning	09318KA2
Advantage GC2 PCR Kit	Takara, Japan	639119
TA cloning with the pUCm-T vector	Sangon, China	B522213