Time origin and structural analysis of the induced CRISPR/cas9 megabase-sized deletions and duplications involving the Cntn6 gene in mice

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Time origin and structural analysis of the induced CRISPR/cas9 megabase-sized deletions and duplications involving the Cntn6 gene in mice. / Pristyazhnyuk, Inna E.; Minina, Julia; Korablev, Alexey et al.

In: Scientific Reports, Vol. 9, No. 1, 14161, 02.10.2019.

Research output: Contribution to journal › Article › peer-review

BibTeX

@article{244cba4f740d438bad2abb577b5645a0,

title = "Time origin and structural analysis of the induced CRISPR/cas9 megabase-sized deletions and duplications involving the Cntn6 gene in mice",

abstract = "In a previous study using one-step CRISPR/Cas9 genome editing in mouse zygotes, we created five founders carrying a 1,137 kb deletion and two founders carrying the same deletion, plus a 2,274 kb duplication involving the Cntn6 gene (encoding contactin-6). Using these mice, the present study had the following aims: (i) to establish stage of origin of these rearrangements; (ii) to determine the fate of the deleted DNA fragments; and (iii) to estimate the scale of unpredicted DNA changes accompanying the rearrangements. The present study demonstrated that all targeted deletions and duplications occurred at the one-cell stage and more often in one pronucleus only. FISH analysis revealed that there were no traces of the deleted DNA fragments either within chromosome 6 or on other chromosomes. These data were consistent with the Southern blot analysis showing that chromosomes with deletion often had close to expected sizes of removed DNA fragments. High-throughput DNA sequencing of two homozygotes for duplication demonstrated that there were no unexpected significant or scale DNA changes either at the gRNA and joint sites or other genome sites. Thus, our data suggested that CRISPR/Cas9 technology could generate megabase-sized deletions and duplications in mouse gametes at a reasonably specific level.",

keywords = "DNA-REPLICATION, GENOME, GENERATION, RECOMBINATION, CRISPR-CAS9, INVERSIONS, MUTATIONS, EFFICIENT, VARIANTS, CELLS",

author = "Pristyazhnyuk, {Inna E.} and Julia Minina and Alexey Korablev and Irina Serova and Veniamin Fishman and Maria Gridina and Rozhdestvensky, {Timofey S.} and Leonid Gubar and Skryabin, {Boris V.} and Serov, {Oleg L.}",

year = "2019",

month = oct,

day = "2",

doi = "10.1038/s41598-019-50649-4",

language = "English",

volume = "9",

journal = "Scientific Reports",

issn = "2045-2322",

publisher = "Nature Publishing Group",

number = "1",

}

RIS

TY - JOUR

T1 - Time origin and structural analysis of the induced CRISPR/cas9 megabase-sized deletions and duplications involving the Cntn6 gene in mice

AU - Pristyazhnyuk, Inna E.

AU - Minina, Julia

AU - Korablev, Alexey

AU - Serova, Irina

AU - Fishman, Veniamin

AU - Gridina, Maria

AU - Rozhdestvensky, Timofey S.

AU - Gubar, Leonid

AU - Skryabin, Boris V.

AU - Serov, Oleg L.

PY - 2019/10/2

Y1 - 2019/10/2

N2 - In a previous study using one-step CRISPR/Cas9 genome editing in mouse zygotes, we created five founders carrying a 1,137 kb deletion and two founders carrying the same deletion, plus a 2,274 kb duplication involving the Cntn6 gene (encoding contactin-6). Using these mice, the present study had the following aims: (i) to establish stage of origin of these rearrangements; (ii) to determine the fate of the deleted DNA fragments; and (iii) to estimate the scale of unpredicted DNA changes accompanying the rearrangements. The present study demonstrated that all targeted deletions and duplications occurred at the one-cell stage and more often in one pronucleus only. FISH analysis revealed that there were no traces of the deleted DNA fragments either within chromosome 6 or on other chromosomes. These data were consistent with the Southern blot analysis showing that chromosomes with deletion often had close to expected sizes of removed DNA fragments. High-throughput DNA sequencing of two homozygotes for duplication demonstrated that there were no unexpected significant or scale DNA changes either at the gRNA and joint sites or other genome sites. Thus, our data suggested that CRISPR/Cas9 technology could generate megabase-sized deletions and duplications in mouse gametes at a reasonably specific level.

AB - In a previous study using one-step CRISPR/Cas9 genome editing in mouse zygotes, we created five founders carrying a 1,137 kb deletion and two founders carrying the same deletion, plus a 2,274 kb duplication involving the Cntn6 gene (encoding contactin-6). Using these mice, the present study had the following aims: (i) to establish stage of origin of these rearrangements; (ii) to determine the fate of the deleted DNA fragments; and (iii) to estimate the scale of unpredicted DNA changes accompanying the rearrangements. The present study demonstrated that all targeted deletions and duplications occurred at the one-cell stage and more often in one pronucleus only. FISH analysis revealed that there were no traces of the deleted DNA fragments either within chromosome 6 or on other chromosomes. These data were consistent with the Southern blot analysis showing that chromosomes with deletion often had close to expected sizes of removed DNA fragments. High-throughput DNA sequencing of two homozygotes for duplication demonstrated that there were no unexpected significant or scale DNA changes either at the gRNA and joint sites or other genome sites. Thus, our data suggested that CRISPR/Cas9 technology could generate megabase-sized deletions and duplications in mouse gametes at a reasonably specific level.

KW - DNA-REPLICATION

KW - GENOME

KW - GENERATION

KW - RECOMBINATION

KW - CRISPR-CAS9

KW - INVERSIONS

KW - MUTATIONS

KW - EFFICIENT

KW - VARIANTS

KW - CELLS

UR - http://www.scopus.com/inward/record.url?scp=85072912398&partnerID=8YFLogxK

U2 - 10.1038/s41598-019-50649-4

DO - 10.1038/s41598-019-50649-4

M3 - Article

C2 - 31578377

AN - SCOPUS:85072912398

VL - 9

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

IS - 1

M1 - 14161

ER -

ID: 21805182