Creation and Analysis of Related Genetic Characteristics of BALB/cA.Cg.SHJH hr Mice

doi:10.12300/j.issn.1674-5817.2023.055

Abstract

Abstract:

Objective To introduce the Hr gene of spontaneously mutated SHJH ^hr mice into BALB/cAShjh inbred mice with clear genetic background,and provide a basis for study on the molecular mechanism of Hr gene mutation-induced abnormal phenotype and the application of this model. Methods Using a backcross-intercross breeding method guided by phenotypic monitoring, mutant genes from SHJH ^hr mice bred by spontaneous mutation were introduced into inbred BALB/cAShjh mice by homozygous mutation introgression, and the mice were bred into BALB/cA.Cg.SHJH ^hr (abbreviated as C.Cg.SHJH ^hr ) mice after 10 generations. The genotypes of 90 single nucleotide polymorphism (SNP) detection sites were analyzed in C.Cg.SHJH ^hr mice by multiplex PCR library construction followed by next generation sequencing. Then 14 biochemical locus marker genes were detected in C.Cg.SHJH ^hr mice according to the method of GB/T 14927.1-2008. Finally, whole genome exon sequencing was utilized to detect the mutated genes in this mouse. Results From May 2018 to March 2022, a total of 10 generations of backcross-intercross were conducted to complete the construction of the C.Cg.SHJH ^hr mouse line. Among the 90 SNPs loci detected, except for rs13484115 and rs13484116, all the other loci had the same genotype as the recipient mice BALB/cAShjh. The results of biochemical marker gene detection showed that all the 14 loci of the mouse were the same as those of the recipient mouse. Whole genome exon sequencing found that the mouse had 109 site mutations compared with the recipient mouse strain, including 71 synonymous mutations, 1 stopgain, 37 missense mutations, and 20 genes involved in protein sequence alterations (including the reported Hr gene). Conclusion C.Cg.SHJH ^hr mice were created. Through exon sequencing and genetic analysis, three Hr mutated genes and associated mutated genes that mainly cause phenotypic variations were identified, which provides a basis for expanding the application of C.Cg.SHJH ^hr mice in biomedical research.

Key words: BALB/cA.Cg.SHJH ^hr mice, Exon sequencing, Mutation, Genetic characteristics

CLC Number:

R-332

Xiaoqian TAN,Hao YANG,Huiqing TANG,et al. Creation and Analysis of Related Genetic Characteristics of BALB/cA.Cg.SHJH ^hr Mice[J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 363-370. DOI: 10.12300/j.issn.1674-5817.2023.055.

Figures/Tables 6

References 24

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位点Loci	C.Cg.SHJH ^hr	BALB/cA
Car2	b	b
Ce2	a	a
Es1	b	b
Es3	a	a
Es10	a	a
Gpd1	b	b
Gpi1	a	a
Hbb	d	d
Idh1	a	a
Mod1	a	a
Akp1	b	b
Pgm1	a	a
Trf	b	b
Pep3	a	a

位点Loci	C.Cg.SHJH ^hr	BALB/cA
Car2	b	b
Ce2	a	a
Es1	b	b
Es3	a	a
Es10	a	a
Gpd1	b	b
Gpi1	a	a
Hbb	d	d
Idh1	a	a
Mod1	a	a
Akp1	b	b
Pgm1	a	a
Trf	b	b
Pep3	a	a

突变位置 Mutation position	碱基变化 Base alteration	基因 Gene	涉及疾病/细胞功能 Related diseases/cell functions	下游信号通路调控 Signal pathway regulation	PolyPhen-2计算得分 PolyPhen-2 scores
70559773	C→T	Hr	全身性脱毛，伴有丘疹性病变的心房颤动	是	1.000
73523427	A→G	Nudet15	造血干细胞和祖细胞急性损失	是	0.000
74744708	T→C	esd	甲醛分解代谢	是	0.505
74811692	T→C	Lrch1	细胞对趋化因子的反应	否	0.047
74813612	T→C
75031916	G→A	5031414D18Rik	自噬体成熟、糖原、脂质代谢过程的调节	否	0.022
75032403	C→A
75036145	A→G
75125178	A→G	Lrrc63	膜蛋白	否	-
75224148	C→T	Lcp1	肌动蛋白丝束组件	否	0.166
78510551	C→G	Akap11	体型缩小，活动能力减退	否	0.997
79401743	G→A	Mtrf1	线粒体翻译终止；耳聋，痉挛性共济失调	否	0.959
79401763	G→T	Mtrf1
79406608	A→G	Mtrf1
79427505	C→G	Kbtbd	蛋白酶体介导的泛素依赖蛋白的分解代谢	是	0.006
79452831	G→C	Kbtbd	蛋白酶体介导的泛素依赖蛋白的分解代谢	是	0.006
79573278	A→G	Elf1	RNA聚合酶Ⅱ转录的正调控	是	0.000
79595387	G→A	Sugt1	参与动粒功能	是	0.000
79637996	C→G	Lect1	内皮细胞增殖负调控	否	0.005
79656659	A→G
79656692	T→C
86966512	G→A	Diaph3	遗传性听神经病变，先天性造血障碍性贫血	是	0.958
96201334	G→T	Klhl1	肌动蛋白结合蛋白	否	0.001
88469236	C→G	Pcdh20	细胞黏附	否	0.000
99046045	G→C	Mzt1	微管蛋白复合体组成	是	-
99061539	G→C	Bora	蛋白激酶活性的激活，细胞周期过程的调控，蛋白质定位的调节	是	0.995
99068023	C→T	Bora
99068337	G→C
99068407	C→G
99068529	A→G
99068542	A→G
99072637	T→A
99072704	G→A
99084109	C→A	Dis3	CUT分解代谢，RNA加工，rRNA分解代谢	是	0.000
99099549	G→T	Dis3	CUT分解代谢，RNA加工，rRNA分解代谢	是	0.000
99101172	A→G	Pibf1	有丝分裂核分裂，细胞因子产生，蛋白质磷酸化	否	0.982

突变位置 Mutation position	碱基变化 Base alteration	基因 Gene	涉及疾病/细胞功能 Related diseases/cell functions	下游信号通路调控 Signal pathway regulation	PolyPhen-2计算得分 PolyPhen-2 scores
70559773	C→T	Hr	全身性脱毛，伴有丘疹性病变的心房颤动	是	1.000
73523427	A→G	Nudet15	造血干细胞和祖细胞急性损失	是	0.000
74744708	T→C	esd	甲醛分解代谢	是	0.505
74811692	T→C	Lrch1	细胞对趋化因子的反应	否	0.047
74813612	T→C
75031916	G→A	5031414D18Rik	自噬体成熟、糖原、脂质代谢过程的调节	否	0.022
75032403	C→A
75036145	A→G
75125178	A→G	Lrrc63	膜蛋白	否	-
75224148	C→T	Lcp1	肌动蛋白丝束组件	否	0.166
78510551	C→G	Akap11	体型缩小，活动能力减退	否	0.997
79401743	G→A	Mtrf1	线粒体翻译终止；耳聋，痉挛性共济失调	否	0.959
79401763	G→T	Mtrf1
79406608	A→G	Mtrf1
79427505	C→G	Kbtbd	蛋白酶体介导的泛素依赖蛋白的分解代谢	是	0.006
79452831	G→C	Kbtbd	蛋白酶体介导的泛素依赖蛋白的分解代谢	是	0.006
79573278	A→G	Elf1	RNA聚合酶Ⅱ转录的正调控	是	0.000
79595387	G→A	Sugt1	参与动粒功能	是	0.000
79637996	C→G	Lect1	内皮细胞增殖负调控	否	0.005
79656659	A→G
79656692	T→C
86966512	G→A	Diaph3	遗传性听神经病变，先天性造血障碍性贫血	是	0.958
96201334	G→T	Klhl1	肌动蛋白结合蛋白	否	0.001
88469236	C→G	Pcdh20	细胞黏附	否	0.000
99046045	G→C	Mzt1	微管蛋白复合体组成	是	-
99061539	G→C	Bora	蛋白激酶活性的激活，细胞周期过程的调控，蛋白质定位的调节	是	0.995
99068023	C→T	Bora
99068337	G→C
99068407	C→G
99068529	A→G
99068542	A→G
99072637	T→A
99072704	G→A
99084109	C→A	Dis3	CUT分解代谢，RNA加工，rRNA分解代谢	是	0.000
99099549	G→T	Dis3	CUT分解代谢，RNA加工，rRNA分解代谢	是	0.000
99101172	A→G	Pibf1	有丝分裂核分裂，细胞因子产生，蛋白质磷酸化	否	0.982

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