| 1. |
- Arlien-Soborg, Mai C., et al.
(författare)
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Acromegaly management in the Nordic countries: A Delphi consensus survey
- 2024
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Ingår i: Clinical Endocrinology. - : WILEY. - 0300-0664 .- 1365-2265.
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Tidskriftsartikel (refereegranskat)abstract
- ObjectiveAcromegaly is associated with increased morbidity and mortality if left untreated. The therapeutic options include surgery, medical treatment, and radiotherapy. Several guidelines and recommendations on treatment algorithms and follow-up exist. However, not all recommendations are strictly evidence-based. To evaluate consensus on the treatment and follow-up of patients with acromegaly in the Nordic countries.MethodsA Delphi process was used to map the landscape of acromegaly management in Denmark, Sweden, Norway, Finland, and Iceland. An expert panel developed 37 statements on the treatment and follow-up of patients with acromegaly. Dedicated endocrinologists (n = 47) from the Nordic countries were invited to rate their extent of agreement with the statements, using a Likert-type scale (1-7). Consensus was defined as >= 80% of panelists rating their agreement as >= 5 or <= 3 on the Likert-type scale.ResultsConsensus was reached in 41% (15/37) of the statements. Panelists agreed that pituitary surgery remains first line treatment. There was general agreement to recommend first-generation somatostatin analog (SSA) treatment after failed surgery and to consider repeat surgery. In addition, there was agreement to recommend combination therapy with first-generation SSA and pegvisomant as second- or third-line treatment. In more than 50% of the statements, consensus was not achieved. Considerable disagreement existed regarding pegvisomant monotherapy, and treatment with pasireotide and dopamine agonists.ConclusionThis consensus exploration study on the management of patients with acromegaly in the Nordic countries revealed a relatively large degree of disagreement among experts, which mirrors the complexity of the disease and the shortage of evidence-based data.
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| 2. |
- Arlien-Søborg, Mai C., et al.
(författare)
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Acromegaly management in the nordic countries : a Delphi consensus survey
- 2024
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Ingår i: Clinical Endocrinology. - : John Wiley & Sons. - 0300-0664 .- 1365-2265.
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Tidskriftsartikel (refereegranskat)abstract
- Objective: Acromegaly is associated with increased morbidity and mortality if left untreated. The therapeutic options include surgery, medical treatment, and radiotherapy. Several guidelines and recommendations on treatment algorithms and follow-up exist. However, not all recommendations are strictly evidence-based. To evaluate consensus on the treatment and follow-up of patients with acromegaly in the Nordic countries.Methods: A Delphi process was used to map the landscape of acromegaly management in Denmark, Sweden, Norway, Finland, and Iceland. An expert panel developed 37 statements on the treatment and follow-up of patients with acromegaly. Dedicated endocrinologists (n = 47) from the Nordic countries were invited to rate their extent of agreement with the statements, using a Likert-type scale (1−7). Consensus was defined as ≥80% of panelists rating their agreement as ≥5 or ≤3 on the Likert-type scale.Results: Consensus was reached in 41% (15/37) of the statements. Panelists agreed that pituitary surgery remains first line treatment. There was general agreement to recommend first-generation somatostatin analog (SSA) treatment after failed surgery and to consider repeat surgery. In addition, there was agreement to recommend combination therapy with first-generation SSA and pegvisomant as second- or third-line treatment. In more than 50% of the statements, consensus was not achieved. Considerable disagreement existed regarding pegvisomant monotherapy, and treatment with pasireotide and dopamine agonists.Conclusion: This consensus exploration study on the management of patients with acromegaly in the Nordic countries revealed a relatively large degree of disagreement among experts, which mirrors the complexity of the disease and the shortage of evidence-based data.
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| 3. |
- Abeid, Muzdalifat, 1973-, et al.
(författare)
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Community perceptions of rape and child sexual abuse : a qualitative study in rural Tanzania
- 2014
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Ingår i: BMC International Health and Human Rights. - 1472-698X. ; 14, s. 23-
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Tidskriftsartikel (refereegranskat)abstract
- Background: Rape of women and children is recognized as a health and human rights issue in Tanzania and internationally. Exploration of the prevailing perceptions in rural areas is needed in order to expand the understanding of sexual violence in the diversity of Tanzania's contexts. The aim of this study therefore was to explore and understand perceptions of rape of women and children at the community level in a rural district in Tanzania with the added objective of exploring those perceptions that may contribute to perpetuating and/or hindering the disclosure of rape incidences. Methods: A qualitative design was employed using focus group discussions with male and female community members including religious leaders, professionals, and other community members. The discussions centered on causes of rape, survivors of rape, help-seeking and reporting, and gathered suggestions on measures for improvement. Six focus group discussions (four of single gender and two of mixed gender) were conducted. The focus group discussions were recorded, transcribed verbatim, and analyzed using manifest qualitative content analysis. Results: The participants perceived rape of women and children to be a frequent and hidden phenomenon. A number of factors were singled out as contributing to rape, such as erosion of social norms, globalization, poverty, vulnerability of children, alcohol/drug abuse and poor parental care. Participants perceived the need for educating the community to raise their knowledge of sexual violence and its consequences, and their roles as preventive agents. Conclusions: In this rural context, social norms reinforce sexual violence against women and children, and hinder them from seeking help from support services. Addressing the identified challenges may promote help-seeking behavior and improve care of survivors of sexual violence, while changes in social and cultural norms are needed for the prevention of sexual violence.
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| 4. |
- Holmström, Margareta, et al.
(författare)
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SWEDISH NATIONAL REGISTRY FOR BLEEDING DISORDERS – A SECOND REPORT
- 2019
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Ingår i: EAHAD 2019.
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Konferensbidrag (refereegranskat)abstract
- Introduction: Hemophilia Care in Sweden is centralized to three different and certified European Hemophilia Care Centers (EHCCs) (Stockholm, Gothenburg and Malmö[f1]). A recent web- based National registry has been set up for patients with bleeding disorders in Sweden. The registry is mainly funded by Swedish authorities. Methods: A multi- professional steering committee is running the registry with representatives from all three centers including physicians, nurses, physiotherapist and also a patient representative. A web- based platform, Real- Q, is used for the registry. Results: By the 31st Dec 2017, a total number of 1030 patients with bleeding disorders were included in the registry, mainly patients with hemophilia A, B and Von Willebrand disease. Data regarding bleedings, treatment modality and type of product, inhibitor status, viral infections are collected. Likewise patient reported outcome measurements (PROM)- such as pain and quality of life[.The number of patients with hemophilia A, B and Von Willebrand disease in 2016 resp 2017 were as follows:Hemophilia A; n = 243 in 2016 and n= 691 in 2017.Hemophilia B: n = 49 in 2016 and n = 191 in 2017.Von Willebrand disease: n = 11 in 2016 and n = 152 in 2017.[LMW1] are registered.[LMW2] are registered on a regular basis? Discussion/Conclusion: The number of patients in the Swedish National Registry for bleeding disorders has increased significantly during the last year; from a total of 308 Dec 31st 2016 to 1030 in Dec 31st 2017. Increasing amount of data will enable further evaluation of treatment data and also joint status, quality of life and bleeding reports.
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| 5. |
- Holmström, Margareta, et al.
(författare)
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Swedish national registry for bleeding disorders - first report
- 2018
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Ingår i: 11th Annual Congress of the European Association for Haemophilia and Allied Disorders 2018, 7–9 February 2018, Madrid, Spain. Haemophilia, 24 (S1). - : Wiley. - 1365-2516.
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Introduction : Hemophilia care in Sweden is centralized to 3 centers localized in Gothenburg, Malmö and Stockholm. All centers are certi-fied as European Hemophilia Comprehensive Care Centers (EHCCs). Recently a web- based Swedish national registry has been established with funding from Swedish authorities. Methods : One of the conclusions from the earlier reports from the Swedish agency for health technology assessment and assessment of social services (SBU) and the Dental and Pharmaceutical benefits agency (TLV) was that a national registry for hemophilia and other bleeding disorders was needed to be able to follow the long- term effects of the disease and treatment strategies. An application was submitted in 2012 to apply for funding from Swedenʹs municipali-ties and count councils (SKL). The registry was validated as an official national registry. A multi- professional steering committee is running the registry with representatives from all 3 hemophilia centers includ-ing physicians, nurses, physiotherapist and a patient representative. Support regarding legal aspects, IT- solutions, statistics and economy is provided by QRC Stockholm. Results : By now, 780 patients with bleeding disorders are included in the Swedish national Registry and data regarding bleedings, treat-ment with factor concentrate, inhibitor status, mutations, viral infections such as hepatitis C and HIV are collected. Patient reported outcome measurements (PROM)- such as pain and quality of life - HJHS and target joints are followed continuously. Discussion/Conclusion : The establishment of a Swedish National Registry enables us to perform national annual reports, have a close follow- up of our patients and perform clinical research. Currently are working on an on- line patient treatment application recording directly into the registry. Data from the registry will be an important tool for further evaluation of the treatment of hemophilia and how it affects the long- term consequences of the disease.
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| 6. |
- Papakokkinou, Eleni, et al.
(författare)
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Excess Morbidity Persists in Patients With Cushing’s Disease During Long-term Remission : A Swedish Nationwide Study
- 2020
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Ingår i: Journal of Clinical Endocrinology and Metabolism. - Washington : Oxford University Press. - 0021-972X .- 1945-7197. ; 105:8, s. 2616-2624
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Tidskriftsartikel (refereegranskat)abstract
- Context: Whether multisystem morbidity in Cushing's disease (CD) remains elevated during long-term remission is still undetermined.Objective: To investigate comorbidities in patients with CD.Design, setting, and patients: A retrospective, nationwide study of patients with CD identified in the Swedish National Patient Register between 1987 and 2013. Individual medical records were reviewed to verify diagnosis and remission status.Main outcomes: Standardized incidence ratios (SIRs) with 95% confidence intervals (CIs) were calculated by using the Swedish general population as reference. Comorbidities were investigated during three different time periods: (i) during the 3 years before diagnosis, (ii) from diagnosis to 1 year after remission, and (iii) during long-term remission.Results: We included 502 patients with confirmed CD, of whom 419 were in remission for a median of 10 (interquartile range 4 to 21) years. SIRs (95% CI) for myocardial infarction (4.4; 1.2 to 11.4), fractures (4.9; 2.7 to 8.3), and deep vein thrombosis (13.8; 3.8 to 35.3) were increased during the 3-year period before diagnosis. From diagnosis until 1 year after remission, SIRs (95% CI were increased for thromboembolism (18.3; 7.9 to 36.0), stroke (4.9; 1.3 to 12.5), and sepsis (13.6; 3.7 to 34.8). SIRs for thromboembolism (4.9; 2.6 to 8.4), stroke (3.1; 1.8 to 4.9), and sepsis (6.0; 3.1 to 10.6) remained increased during long-term remission.Conclusion: Patients with CD have an increased incidence of stroke, thromboembolism, and sepsis even after remission, emphasizing the importance of early identification and management of risk factors for these comorbidities during long-term follow-up.
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| 7. |
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| 8. |
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| 9. |
- Ragnarsson, Oskar, 1971, et al.
(författare)
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Overall and Disease-Specific Mortality in Patients With Cushing Disease: A Swedish Nationwide Study
- 2019
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Ingår i: Journal of Clinical Endocrinology and Metabolism. - : ENDOCRINE SOC. - 0021-972X .- 1945-7197. ; 104:6, s. 2375-2384
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Tidskriftsartikel (refereegranskat)abstract
- Context: Whether patients with Cushing disease (CD) in remission have increased mortality is still debatable. Objective: To study overall and disease-specific mortality and predictive factors in an unselected nationwide cohort of patients with CD. Design, Patients, and Methods: A retrospective study of patients diagnosed with CD, identified in the Swedish National Patient Registry between 1987 and 2013. Medical records were systematically reviewed to verify the diagnosis. Standardized mortality ratios (SMRs) with 95% CIs were calculated and Cox regression models were used to identify predictors of mortality. Results: Of 502 identified patients with CD (n = 387 women; 77%), 419 (83%) were confirmed to be in remission. Mean age at diagnosis was 43 (SD, 16) years and median follow-up was 13 (interquartile range, 6 to 23) years. The observed number of deaths was 133 vs 54 expected, resulting in an overall SMR of 2.5 (95% CI, 2.1 to 2.9). The commonest cause of death was cardiovascular diseases (SMR, 3.3; 95% CI, 2.6 to 4.3). Excess mortality was also found associated with infections and suicide. For patients in remission, the SMR was 1.9 (95% CI, 1.5 to 2.3); bilateral adrenalectomy and glucocorticoid replacement therapy were independently associated with increased mortality, whereas GH replacement was associated with improved outcome. Conclusion: Findings from this large nationwide study indicate that patients with CD have excess mortality. The findings illustrate the importance of achieving remission and continued active surveillance, along with adequate hormone replacement and evaluation of cardiovascular risk and mental health.
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| 10. |
- Ragnarsson, Oskar, 1971, et al.
(författare)
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The incidence of Cushing’s disease : a nationwide Swedish study
- 2019
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Ingår i: Pituitary. - : Springer. - 1386-341X .- 1573-7403. ; 22:2, s. 179-186
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Tidskriftsartikel (refereegranskat)abstract
- Background: Studies on the incidence of Cushing’s disease (CD) are few and usually limited by a small number of patients. The aim of this study was to assess the annual incidence in a nationwide cohort of patients with presumed CD in Sweden.Methods: Patients registered with a diagnostic code for Cushing’s syndrome (CS) or CD, between 1987 and 2013 were identified in the Swedish National Patient Registry. The CD diagnosis was validated by reviewing clinical, biochemical, imaging, and histopathological data.Results: Of 1317 patients identified, 534 (41%) had confirmed CD. One-hundred-and-fifty-six (12%) patients had other forms of CS, 41 (3%) had probable but unconfirmed CD, and 334 (25%) had diagnoses unrelated to CS. The mean (95% confidence interval) annual incidence between 1987 and 2013 of confirmed CD was 1.6 (1.4–1.8) cases per million. 1987–1995, 1996–2004, and 2005–2013, the mean annual incidence was 1.5 (1.1–1.8), 1.4 (1.0–1.7) and 2.0 (1.7–2.3) cases per million, respectively. During the last time period the incidence was higher than during the first and second time periods (P < 0.05).Conclusion: The incidence of CD in Sweden (1.6 cases per million) is in agreement with most previous reports. A higher incidence between 2005 and 2013 compared to 1987–2004 was noticed. Whether this reflects a truly increased incidence of the disease, or simply an increased awareness, earlier recognition, and earlier diagnosis can, however, not be answered. This study also illustrates the importance of validation of the diagnosis of CD in epidemiological research.
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| 11. |
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| 12. |
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| 13. |
- Springett, Jane, 1952-, et al.
(författare)
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Närsjukvård : bakgrund, erfarenheter och pilotstudie
- 2005
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Rapport (övrigt vetenskapligt/konstnärligt)abstract
- Närsjukvård är ett centralt begrepp i ett förändringsarbete som för genomförs i nordöstra Skåne för att utveckla hälso- och sjukvårdsväsendet. Det ingår därmed som en av de centrala delarna av Region Skånes vision om hälso- och sjukvård: Skånsk livskraft – vård och hälsa. Syftet med denna rapport är att ge en bakgrund till begreppet (Del A) och att presentera en del preliminära rön beträffande hur olika aktörer i nordöstra Skåne uppfattar begreppet (Del B). Del A ger en översikt kring ursprunget till begreppet Närsjukvård inom ramen för de förändringar i hälso- och sjukvården som sker i Sverige i stort. Den beskriver sedan vilka slags förändringar som har planerats på politisk nivå och som nu håller på att genomföras under detta paraplybegrepp, nationellt, regionalt och lokalt. För detta syfte används statliga dokument och publicerade utvärderingsstudier i stor utsträckning som källmaterial. Denna del ska därför inte ses som en heltäckande översikt. Del B inriktas på att belysa hur långt förverkligandet av idén om Närsjukvård har kommit inom regionen. Avsnittet är en kartläggning av olika aktörers förståelse av Närsjukvård i den nordöstra delen av Region Skåne. Forskningsfrågorna inriktades på hur folk pratade om Närsjukvård, det vill säga på hur de förstod och använde begreppet.
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| 14. |
- Springett, Jane, et al.
(författare)
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Närsjukvård : bakgrund, erfarenheter och pilotstudie
- 2005
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Bok (övrigt vetenskapligt/konstnärligt)abstract
- Närsjukvård är ett centralt begrepp i ett förändringsarbete som för genomförs i nordöstra Skåne för att utveckla hälso- och sjukvårdsväsendet. Det ingår därmed som en av de centrala delarna av Region Skånes vision om hälso- och sjukvård: Skånsk livskraft – vård och hälsa. Syftet med denna rapport är att ge en bakgrund till begreppet (Del A) och att presentera en del preliminära rön beträffande hur olika aktörer i nordöstra Skåne uppfattar begreppet (Del B). Del A ger en översikt kring ursprunget till begreppet Närsjukvård inom ramen för de förändringar i hälso- och sjukvården som sker i Sverige i stort. Den beskriver sedan vilka slags förändringar som har planerats på politisk nivå och som nu håller på att genomföras under detta paraplybegrepp, nationellt, regionalt och lokalt. För detta syfte används statliga dokument och publicerade utvärderingsstudier i stor utsträckning som källmaterial. Denna del ska därför inte ses som en heltäckande översikt. Del B inriktas på att belysa hur långt förverkligandet av idén om Närsjukvård har kommit inom regionen. Avsnittet är en kartläggning av olika aktörers förståelse av Närsjukvård i den nordöstra delen av Region Skåne. Forskningsfrågorna inriktades på hur folk pratade om Närsjukvård, det vill säga på hur de förstod och använde begreppet.
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| 15. |
- Aarnio, Pauliina, et al.
(författare)
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Husband's role in handling pregnancy complications in Mangochi District, Malawi : A call for increased focus on community level male involvement
- 2018
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Ingår i: Sexual & Reproductive HealthCare. - : ELSEVIER IRELAND LTD. - 1877-5756 .- 1877-5764. ; 16, s. 61-66
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Tidskriftsartikel (refereegranskat)abstract
- Objective: The objective of the current study is to provide information about husbands' role in decision-making and healthcare seeking in cases of pregnancy complications in Mangochi district, Malawi with an analysis of qualitative interviews using the concepts of "capital" and "field" from Bourdieu's social field theory. Study design: Twelve husbands and wives who had experienced pregnancy complications and six key informants from a semi-rural area of Mangochi district were interviewed individually. Thematic analysis was conducted based on the concepts of capital and field in Bourdieu's social field theory. Results: Husbands have significant economic and symbolic capital in decisions about healthcare seeking during instances of pregnancy complications as a result of their roles as father, head of the household and main income earner. Lack of money is the only acceptable reason for husbands to deny their wives healthcare. Husbands have limited access to knowledge of maternal health, which can compromise their decisions about seeking healthcare. Joint decision-making within families can be bypassed to allow for prompt healthcare seeking in emergencies. Conclusions: Husbands are important decision makers regarding seeking healthcare for pregnancy complications because of their economic and symbolic power and despite their limited access to knowledge of maternal health. Maternal healthcare seeking practices would benefit from wives gaining an empowered role as well as improved knowledge of maternal health among husbands.
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| 16. |
- Aarnio, Pauliina, et al.
(författare)
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Male involvement in antenatal HIV counseling and testing : exploring men's perceptions in rural Malawi
- 2009
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Ingår i: AIDS Care. - : Informa UK Limited. - 0954-0121 .- 1360-0451. ; 21:12, s. 1537-1546
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Tidskriftsartikel (refereegranskat)abstract
- Antenatal care can act as an excellent tool to improve access to HIV counseling and testing services. This paper investigates an issue that may weaken its potential, namely lack of male involvement. We explored married men's perceptions of HIV in pregnancy and male involvement in antenatal HIV testing and counseling in Southern Malawi through 11 focus group discussions and a cross-sectional survey (n=388). The main findings were that men were largely unaware of available antenatal HIV testing and counseling services, and perceived it overall problematic to attend female-oriented health care. Most men supported provision of antenatal HIV testing. They perceived husbands to participate in the process indirectly through spousal communication, being faithful during pregnancy, and supporting the wife if found HIV-positive. Involvement of husbands was compromised by men's reluctance to learn their HIV status and the threat that HIV poses on marriage. Men stressed the importance of prior spousal agreement of antenatal HIV testing and considered HIV testing without their consent a valid reason for divorce. We suggest that male involvement in antenatal HIV testing requires refocusing of information and health services to include men. To avoid negative social outcomes for women, comprehensive and early involvement of men is essential.
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| 17. |
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| 18. |
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| 19. |
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| 20. |
- Adelmann, Kenneth, et al.
(författare)
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Språkutveckling, medier och demokrati
- 2014
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Ingår i: Medie- och informationskunnighet i Norden. - Göteborg : Nordicom. - 9789186523886 ; , s. 117-129
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Bokkapitel (övrigt vetenskapligt/konstnärligt)
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| 21. |
- Adelmann, Kent, et al.
(författare)
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Språkutveckling, medier och demokrati
- 2014
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Ingår i: Medie- och informationskunnighet i Norden. - : Nordicom. - 9789186523886 ; , s. 117-121
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Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- Vi som har skrivit denna artikel är alla verksamma inom forskningsmiljön Svenska med didaktisk inriktning (SMDI) vid Malmö högskola. Frågor om språkutveckling i bred bemärkelse står här i centrum. För närvarande är vi engagerade i projektet ”SMDI och lärande i medielandskapet 2.0”. Vår teoretiska plattform kan beskrivas som medieekologisk, vilket kortfattat uttryckt innebär att vi är intresserade av de mångfaldiga och komplexa relationerna mellan medier och kommunikativa kompetenser (Elmfeldt & Erixon 2007; Erixon 2012; Hayles 2002). Dessa relationer förstås därför inte, som så ofta annars i skolsammanhang, i termer av enkelriktad påverkan eller effekter (exempelvis datorns och internets negativa inverkan på skriftspråket). Vår huvudpoäng i denna artikel är att medie- och informationskunnighet, MIK, handlar om, borde handla om, framför allt två saker: språkutveckling och demokrati.
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| 22. |
- Al-Shamkhi, Nasrin, 1985-, et al.
(författare)
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Pituitary function before and after surgery for nonfunctioning pituitary adenomas-data from the Swedish Pituitary Register.
- 2023
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Ingår i: European journal of endocrinology. - : Bioscientifica. - 1479-683X .- 0804-4643. ; 189:2, s. 217-224
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Tidskriftsartikel (refereegranskat)abstract
- Data on pre- and postoperative pituitary function in nonfunctioning pituitary adenomas (NFPA) are not consistent. We aimed to investigate pituitary function before and up to 5 years after transsphenoidal surgery with emphasis on the hypothalamic-pituitary-adrenal axis (HPA).Data from the Swedish Pituitary Register was used to analyze anterior pituitary function in 838 patients with NFPA diagnosed between 1991 and 2014. Patients who were reoperated or had received radiotherapy were excluded.Preoperative ACTH, TSH, LH/FSH, and GH deficiencies were reported in 31% (236/755), 39% (300/769), 51% (378/742), and 28% (170/604) of the patients, respectively. Preoperative median tumor volume was 5.0 (2.4-9.0) cm3. Among patients with preoperative, 1 year and 5 years postoperative data on the HPA axis (n = 428), 125 (29%) were ACTH-deficient preoperatively. One year postoperatively, 26% (32/125) of them had recovered ACTH function while 23% (70/303) patients had developed new ACTH deficiency. Thus, 1 year postoperatively, 163 (38%) patients were ACTH-deficient (P < .001 vs. preoperatively). No further increase was seen 5 years postoperatively (36%, P = .096). At 1 year postoperatively, recoveries in the TSH and LH/FSH axes were reported in 14% (33/241) and 15% (46/310), respectively, and new deficiencies in 22% (88/403) and 29% (83/288), respectively.Adrenocorticotrophic hormone deficiency increased significantly at 1 year postoperatively. Even though not significant, some patients recovered from or developed new deficiency between 1 and 5 years postoperatively. This pattern was seen in all axes. Our study emphasizes that continuous individual evaluations are needed during longer follow-up of patients operated for NFPA.
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| 23. |
- Andersson, Mariette, et al.
(författare)
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Genome editing in potato via CRISPR-Cas9 ribonucleoprotein delivery
- 2018
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Ingår i: Physiologia Plantarum. - : Wiley. - 0031-9317 .- 1399-3054. ; 164, s. 378-384
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Tidskriftsartikel (refereegranskat)abstract
- Clustered regularly interspaced short palindromic repeats and CRISPR-associated protein-9 (CRISPR-Cas9) can be used as an efficient tool for genome editing in potato (Solanum tuberosum). From both a scientific and a regulatory perspective, it is beneficial if integration of DNA in the potato genome is avoided. We have implemented a DNA-free genome editing method, using delivery of CRISPR-Cas9 ribonucleoproteins (RNPs) to potato protoplasts, by targeting the gene encoding a granule bound starch synthase (GBSS, EC 2.4.1.242). The RNP method was directly implemented using previously developed protoplast isolation, transfection and regeneration protocols without further adjustments. Cas9 protein was preassembled with RNA produced either synthetically or by in vitro transcription. RNP with synthetically produced RNA (cr-RNP) induced mutations, i.e. indels, at a frequency of up to 9%, with all mutated lines being transgene-free. A mutagenesis frequency of 25% of all regenerated shoots was found when using RNP with in vitro transcriptionally produced RNA (IVT-RNP). However, more than 80% of the shoots with confirmed mutations had unintended inserts in the cut site, which was in the same range as when using DNA delivery. The inserts originated both from DNA template remnants from the in vitro transcription, and from chromosomal potato DNA. In 2-3% of the regenerated shoots from the RNP-experiments, mutations were induced in all four alleles resulting in a complete knockout of the GBSS enzyme function.
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| 25. |
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