NDLI: The genomes of precision edited cloned calves show no evidence for off-target events or increased de novo mutagenesis

Content Provider	Springer Nature : BioMed Central
Author	Jivanji, Swati Harland, Chad Cole, Sally Brophy, Brigid Garrick, Dorian Snell, Russell Littlejohn, Mathew Laible, Götz
Abstract	Background Animal health and welfare are at the forefront of public concern and the agricultural sector is responding by prioritising the selection of welfare-relevant traits in their breeding schemes. In some cases, welfare-enhancing traits such as horn-status (i.e., polled) or diluted coat colour, which could enhance heat tolerance, may not segregate in breeds of primary interest, highlighting gene-editing tools such as the CRISPR-Cas9 technology as an approach to rapidly introduce variation into these populations. A major limitation preventing the acceptance of CRISPR-Cas9 mediated gene-editing, however, is the potential for off-target mutagenesis, which has raised concerns about the safety and ultimate applicability of this technology. Here, we present a clone-based study design that has allowed a detailed investigation of off-target and de novo mutagenesis in a cattle line bearing edits in the PMEL gene for diluted coat-colour. Results No off-target events were detected from high depth whole genome sequencing performed in precursor cell-lines and resultant calves cloned from those edited and non-edited cell lines. Long molecule sequencing at the edited site and plasmid-specific PCRs did not reveal structural variations and/or plasmid integration events in edited samples. Furthermore, an in-depth analysis of de novo mutations across the edited and non-edited cloned calves revealed that the mutation frequency and spectra were unaffected by editing status. Cells in culture, however, appeared to have a distinct mutation signature where de novo mutations were predominantly C > A mutations, and in cloned calves they were predominantly T > G mutations, deviating from the expected excess of C > T mutations. Conclusions We found no detectable CRISPR-Cas9 associated off-target mutations in the gene-edited cells or calves derived from the gene-edited cell line. Comparison of de novo mutation in two gene-edited calves and three non-edited control calves did not reveal a higher mutation load in any one group, gene-edited or control, beyond those anticipated from spontaneous mutagenesis. Cell culture and somatic cell nuclear transfer cloning processes contributed the major source of contrast in mutational profile between samples.
Related Links	https://bmcgenomics.biomedcentral.com/counter/pdf/10.1186/s12864-021-07804-x.pdf
Ending Page	14
Page Count	14
Starting Page	1
File Format	HTM / HTML
ISSN	14712164
DOI	10.1186/s12864-021-07804-x
Journal	BMC Genomics
Issue Number	1
Volume Number	22
Language	English
Publisher	BioMed Central
Publisher Date	2021-06-17
Access Restriction	Open
Subject Keyword	Life Sciences Microarrays Proteomics Animal Genetics and Genomics Microbial Genetics and Genomics Plant Genetics and Genomics
Content Type	Text
Resource Type	Article
Subject	Biotechnology Genetics
Journal Impact Factor	3.5/2023
5-Year Journal Impact Factor	4.1/2023

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

The genomes of precision edited cloned calves show no evidence for off-target events or increased de novo mutagenesis

Low incidence of SNVs and indels in trio genomes of Cas9-mediated multiplex edited sheep

De novo copy number variations in cloned dogs from the same nuclear donor

Correction to: L_RNA_scaffolder: scaffolding genomes with transcripts

OffScan: a universal and fast CRISPR off-target sites detection tool

HGA: de novo genome assembly method for bacterial genomes using high coverage short sequencing reads

TREC-IN: gene knock-in genetic tool for genomes cloned in yeast

The distribution and mutagenesis of short coding INDELs from 1,128 whole exomes

Evaluating the quality of the 1000 genomes project data

The genomes of precision edited cloned calves show no evidence for off-target events or increased de novo mutagenesis

Similar Documents

The genomes of precision edited cloned calves show no evidence for off-target events or increased de novo mutagenesis

Low incidence of SNVs and indels in trio genomes of Cas9-mediated multiplex edited sheep

De novo copy number variations in cloned dogs from the same nuclear donor

Correction to: L_RNA_scaffolder: scaffolding genomes with transcripts

OffScan: a universal and fast CRISPR off-target sites detection tool

HGA: de novo genome assembly method for bacterial genomes using high coverage short sequencing reads

TREC-IN: gene knock-in genetic tool for genomes cloned in yeast

The distribution and mutagenesis of short coding INDELs from 1,128 whole exomes

Evaluating the quality of the 1000 genomes project data

The genomes of precision edited cloned calves show no evidence for off-target events or increased de novo mutagenesis