CRISPR/Cas9 technology allows accurate, marker-less genome editing. We report a detailed, robust, and streamlined protocol for CRISPR/Cas9 genome editing in , based on the widely used MoClo-Yeast Toolkit (https://www.addgene.org/kits/moclo-ytk/). This step-by-step protocol guides the reader from sgRNA design to verification of the desired genome editing event and provides preassembled plasmids for cloning the sgRNA(s), making this technology easily accessible to any yeast research group. For complete details on the use and execution of this protocol, please refer to Novarina et al. (2021).
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9192979 | PMC |
| http://dx.doi.org/10.1016/j.xpro.2022.101358 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
A versatile site-directed gene trap strategy to manipulate gene activity and control gene expression in Caenorhabditis elegans.
PLoS Genet
January 2025
Department of Integrative Biology, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
The ability to manipulate gene activity and control transgene expression is essential to study gene function. While several genetic tools for modifying genes or controlling expression separately are available for Caenorhabditis elegans, there are no genetic approaches to generate mutations that simultaneously disrupt gene function and provide genetic access to the cells expressing the disrupted gene. To achieve this, we developed a versatile gene trap strategy based on cGAL, a GAL4-UAS bipartite expression system for C.
View Article and Find Full Text PDFTransgene-free genome editing in poplar.
New Phytol
January 2025
Department of Plant Biotechnology and Bioinformatics, Ghent University, Ghent, 9052, Belgium.
Precise gene-editing methods are valuable tools to enhance genetic traits. Gene editing is commonly achieved via stable integration of a gene-editing cassette in the plant's genome. However, this technique is unfavorable for field applications, especially in vegetatively propagated plants, such as many commercial tree species, where the gene-editing cassette cannot be segregated away without breaking the genetic constitution of the elite variety.
View Article and Find Full Text PDFTransthyretin Cardiac Amyloidosis: Current and Emerging Therapies.
Curr Cardiol Rep
January 2025
The Pauley Heart Center, Virginia Commonwealth University, 1200 East Broad Street West Hospital, 8th Floor, West Wing, Richmond, VA, 23231, USA.
Purpose Of Review: In this article, we describe current and newer TTR stabilizers, TTR silencers which include small interfering RNA agents (siRNA), antisense oligonucleotides (ASO) and CRISPR-Cas9 gene editing, and TTR depleters, which investigates the use of monoclonal antibodies to remove amyloid fibril deposits for patients with advanced disease.
Recent Findings: Once thought to be a rare and fatal condition, increased recognition, improved non-invasive diagnostic tools, and the explosive development of novel therapies, has transformed the landscape of transthyretin amyloid cardiomyopathy (ATTR-CM). Advances in cardiac imaging with respect to echocardiography, cardiac magnetic resonance imaging (CMR), and radionuclide bone scintigraphy has increased the diagnosis of ATTR-CM over the last twenty years.
The role of nanoparticles in transforming plant genetic engineering: advancements, challenges and future prospects.
Funct Integr Genomics
January 2025
Department of Botany, Maharshi Dayanand University, Rohtak, 124001, India.
Despite years of progress in biotechnology, altering the genetic makeup of many plant species, especially their plastids, remains challenging. The existence of a cell wall poses a significant obstacle to the effectual transportation of biomolecules. Developing efficient methods to introduce genes into plant cells and organelles without causing harm is an ongoing area of research.
View Article and Find Full Text PDFPast and future of cytoplasmic male sterility and heterosis breeding in crop plants.
Plant Cell Rep
January 2025
ICAR-Indian Institute of Pulses Research, Kanpur, Uttar Pradesh, 208024, India.
Plant breeding needs to embrace genetic innovations to ensure stability in crop yields under fluctuating climatic conditions. Development of commercial hybrid varieties has proven to be a sustainable and economical alternative to deliver superior yield, quality and resistance with uniformity in a number of food crops. Cytoplasmic male sterility (CMS), a maternally inherited inability to produce functional pollen, facilitates a three-line system for efficient hybrid seed production strategies in crops.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!