Recent developments in CRISPR/Cas9 genome editing research for edible fungiculture.

Edible fungi are not only delicious but are also rich in nutritional and medicinal value, which is highly sought after by consumers. As the edible fungi industry continues to rapidly advance worldwide, particularly in China, the cultivation of superior and innovative edible fungi strains has become increasingly pivotal. Nevertheless, conventional breeding techniques for edible fungi can be arduous and time-consuming. CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9) is a powerful tool for molecular breeding due to its ability to mediate high-efficiency and high-precision genome modification, which has been successfully applied to many kinds of edible fungi. In this review, we briefly summarized the working mechanism of the CRISPR/Cas9 system and highlighted the application progress of CRISPR/Cas9-mediated genome-editing technology in edible fungi, including Agaricus bisporus, Ganoderma lucidum, Flammulina filiformis, Ustilago maydis, Pleurotus eryngii, Pleurotus ostreatus, Coprinopsis cinerea, Schizophyllum commune, Cordyceps militaris, and Shiraia bambusicola. Additionally, we discussed the limitations and challenges encountered using CRISPR/Cas9 technology in edible fungi and provided potential solutions. Finally, the applications of CRISPR/Cas9 system for molecular breeding of edible fungi in the future are explored.

Ganoderma lucidum is a well-known traditional medicinal mushroom that has attracted considerable attention due to its potential as a promising cell factory for producing high-value bioactive compounds. However, conventional methods for the genetic manipulation of G. lucidum are often time-consuming and labor-intensive, hindering research into the biosynthesis and regulatory mechanisms of its valuable natural products. In recent years, the clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9 (CRISPR/Cas9) system has emerged as a powerful genome editing tool, extensively utilized in life sciences research due to its high efficiency and user-friendliness. This review provides a structured overview of advancements in CRISPR/Cas9-mediated genome editing technology in G. lucidum. We first discuss the development and optimization of the CRISPR system, focusing on the various expression strategies for Cas9 and guide RNA established in G. lucidum. Furthermore, we highlight the application of this system for targeted gene deletion, insertion, and replacement in genome editing, as well as its use in the functional analysis of genes in G. lucidum. In addition, we discuss the limitations and challenges associated with employing CRISPR/Cas9 tools in G. lucidum and provide an outlook on the future development of the CRISPR/Cas9 system and its applications in this organism.

First introduced into mammalian organisms in 2013, the RNA-guided genome editing tool CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9) offers several advantages over conventional ones, such as simple-to-design,...

Cancer is the second leading cause of death globally. Series of sequential, repeated genetic changes and epigenetic modifications are leading to the formation of tumors. These tumors subsequently causing the infected cells to invade and transform their surrounding cells by metastasis are some hallmarks in cancer. Although tremendous efforts have been extended for structurally characterizing the numerous genomic mutations undergoing in cancer cells, there is a lack of information regarding the functions of many mutated genes. Clustered Regularly Interspaced Short Palindromic repeats/CRISPR-associated nuclease 9 (CRISPR/Cas9) has become a robust method for building changes in genome of many organisms. Recent reports have suggested that modification of CRISPR/Cas9 can provide plot form to probe the mechanisms in tumorigenesis and in cancer therapies. This review focuses on the historical perspectives of CRISPR/Cas9. The study highlights the applications and also role in cancer cell genome editing, which is helpful to understand the dynamics. Intense research in progress on mechanism of action of CRISPR/Cas9 has been reviewed and critically discussed. Further, relevant literature on animal models focusing on various approaches has been highlighted to emphasize the therapeutics of CRISPR/Cas9 with current trends and future challenges.

CRISPR/Cas9 and CAR-T cell, collaboration of two revolutionary technologies in cancer immunotherapy, an instruction for successful cancer treatment

Oyster Mushroom (Pleurotus ostreatus)

CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-CRISPR associated nuclease 9) systems have emerged as versatile and convenient (epi)genome editing tools and have become an important player in medical genetic research....

Levels of cadmium (Cd), arsenic (As), mercury (Hg), lead (Pb), iron (Fe), and zinc (Zn) were investigated in 285 samples of 9 species of edible fungi (Lentinus edodes, Auricularia auricula, Pleurotus ostreatus, Tremella fuciformis, Flammulina velutipes, Agrocybe chaxinggu, Armillaria mellea, Agaricus bisporus, and Pholiota nameko), which were collected from markets in Beijing, China. In addition, edible fungi and culture substrates were collected from 7 cultivation bases to examine the role of the substrate in trace metal accumulation. Trace metal concentrations were determined on a dry weight basis. Data showed that all the edible fungi contained trace metals, the levels of which varied among species, and there were significant positive correlations between trace metal (Cd, Pb, and As) concentrations in mushrooms and their substrates. The concentrations of Cd, As, Hg, Pb, Fe, and Zn in the tested fungi ranged from 0.005 to 13.8 mg/kg, nd to 1.62 mg/kg, nd to 0.506 mg/kg, 0.011 to 22.1 mg/kg, 46.3 to 2514 mg/kg, and 14.6 to 289 mg/kg, respectively. In general, concentrations of Cd, As, Hg, Pb, Fe, and Zn were relatively high in L. edodes, whereas Tremella fuciformis and P. nameko had relatively low levels of trace metals. Furthermore, the estimated weekly intake of trace metals was calculated and compared with the WHO/FAO provisional tolerable weekly intake. The estimated weekly intake of Cd, As, Hg, Pb, Fe, and Zn from consuming edible fungi was lower than the provisional tolerable weekly intake. This paper reports information about trace metal concentrations in edible fungi collected from Beijing, China. This paper provides useful information for consumers and regulators about levels of trace metals in edible fungi.

Benefit-risk assessment of metal bioavailability in edible fungi by biomimetic whole digestive tracts with digestion, metabolism, and absorption functions

Introduction: Despite many edible fungi being claimed to be potential sources of functional food, clinical trials on the effect of fungal-based essence on cognitive function are generally lacking. This study aimed to determine the effect of a commercialised fungal-based essence on general health, perceived stress, and fatigue levels, derived from their questionnaire-based perceptions after the consumption of fungi extract. Materials and methods: A randomised, double-blind, placebo-controlled clinical trial was conducted among the healthy Malaysian adult volunteers using the hot water extraction of five edible fungi species, i.e., Boletus edulis, Cordyceps militaris, Ganoderma lucidum, Hericium erinaceus, and Lentinula edodes. Forty participants were allocated a 3:1 ratio to receive fungal-based essence and chicken essence. The questionnaire was carried out at baseline and on day 14, with data from 40 participants being analysed, while the 10-person control group was given chicken essence. Results: The T-test analysis showed that the ingestion of fungal-based essence improved general health, lowered perceived stress, and reduced fatigue levels significantly in both the test and control groups. Conclusion: Our findings support the consistent uptake of fungal-based essence, which could be effective for the improvement of general health and overcoming stress, as well as a promising candidate for use as an anti-fatigue food.

The development of applications for the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9) system has increased greatly in recent years, especially in the area of gene therapy by efficient in vivo genome editing. Although great success has been achieved in repairing and rewriting genomes through homology-directed repair coupled with Cas9 nuclease cleavage, its in vivo efficiency is insufficient for gene therapy. Base editing is a next-generation genome-editing tool that does not involve double-stranded DNA breaks and uses components of the CRISPR system together with other enzymes to make point mutations directly in cellular DNA or RNA. Base editors, composed of an engineered deaminase and a catalytically impaired CRISPR/Cas9 variant, are powerful tools for targeted base editing in cells and organisms. In non-dividing cells, base editors can directly transform one base or base pair into another, efficiently installing a point mutation. Undesired by-products of editing are seldom generated during this procedure. Herein we review the different base-editing platforms, including their deaminase recruitment strategies and editing outcomes, and the in vivo delivery of base editors. Additionally, we summarize therapeutic applications of base editing in disorders of the inner ear.

A comparative assessment of three cultivated edible mushroom species namely Bottom mushroom (Agaricus bisporus), Pearl Oyster (Pleurotus ostreatus), Shiitake Mushroom (Lentinul aedodes) on the cumulative fatty acid compositions through fatty acids methyl esters (FAMEs) was devised. About Thirty-six fatty acids were distinguished in the selected mushroom species to interpret descriptive and analytical information by deriving all free fatty acids to their corresponding FAMEs utilizing gas chromatography-mass spectrometry (GC-MS/MS). Cis-linoleic acid was the predominant fatty acid in the fruit bodies of all studied mushrooms. The range of cis-linoleic acid content in the species was 40.33% to 65.59%. Cis-oleic and heptadesenoic acids, respectively, were the other two main fatty acids. More than 32 fatty acids were found and quantified in addition to the three primary fatty acids that are already mentioned. The sequences of Polyunsaturated fatty acids (PUFA), monounsaturated fatty acids (MUFA) and saturated fatty acids (SFA) within all saturated and unsaturated fatty acids for three mushroom species were PUFA > MUFA > SFA for Agaricus bisporus and Lentinula edodes while the sequence was PUFA > SFA > MUFA for Pleurotus ostreatus. Lentinula edodes presented the highest fatty acids amounts which were followed by Pleurotus ostreatus and Agaricus bisporus respectively. The fatty acid composition of the mushrooms revealed that the unsaturated fatty acids are present in higher amounts than the saturated fatty acids in the analyzed mushrooms.

Gene therapy is based on the principle of the genetic manipulation of DNA or RNA for treating and preventing human diseases. The clustered regularly interspaced short palindromic repeats/CRISPR associated nuclease9 (CRISPR/Cas9) system, derived from the acquired immune system in bacteria and archaea, has provided a new tool for accurate manipulation of genomic sequence to attain a therapeutic result. The advantage of CRISPR which made it an easy and flexible tool for diverse genome editing purposes is that a single protein (Cas9) complex with 2 short RNA sequences, function as a site-specific endonuclease. Recently, application of CRISPR/Cas9 system has become popular for therapeutic aims such as gene therapy. In this article, we review the fundamental mechanisms of CRISPR-Cas9 function and summarize preclinical CRISPR-mediated gene therapy reports on a wide variety of disorders.

CRISPR/Cas9-based tools for targeted genome editing and replication control of HBV.

Clustered regularly interspaced short palindromic repeats/CRISPR associated nuclease 9 (CRISPR-Cas9) system is a rapid technology for gene editing. CRISPR-Cas9 is an RNA guided gene editing tool where Cas9 acts as endonuclease by cutting the target DNA strand. Double Stranded Breaks (DBS) can be repaired by non-homologous end joining (NHEJ) and homology-directed repair (HDR). The NHEJ employs DNA ligase IV to rejoin the broken ends which cause insertion or deletion mutations, whereas HDR repairs the DSBs based on a homologous complementary template and results in perfect repair of broken ends. CRISPR-Cas9 impart diverse advantageous features in contrast with the conventional methods. In this review article, we have discussed CRISPR-Cas9 based genome editing along with its mechanism of action and role in crop improvement.