身為一個熱愛美食、喜歡在城市裡挖掘驚喜的人，臺中公益路一直是我最常出沒的地方之一。這條路可說是「臺中人的美食戰場」，從精緻西餐到創意火鍋，從日式丼飯到義式早午餐，每走幾步，就會有完全不同的特色料理餐廳。

這次我特別花了一整個月，實際造訪了公益路上十間口碑不錯的餐廳。有的是網友熱推的打卡名店，也有隱藏在巷弄裡的小驚喜。我以環境氛圍、口味表現、價格CP值與再訪意願為基準，整理出這篇實測評比。希望能幫正在猶豫去哪裡吃飯的你，找到那一間「吃完會想再來」的餐廳。

評比標準與整理方向

這次我走訪的10家餐廳橫跨不同料理類型，從高質感牛排館到巷弄系早午餐，每一間都有自己獨特的風格。為了讓整體比較更客觀，我依照以下四大面向進行評比，並搭配實際用餐體驗來打分。

整體而言，我希望這份評比不只是「哪家好吃」，而是幫你在不同情境下（約會、家庭聚餐、朋友小聚、商業午餐）都能快速找到合適的選擇。畢竟，美食不只是味覺的滿足，更是一段段與朋友共享的生活記憶。

10間臺中公益路餐廳評比懶人包

公益路向來是臺中人聚餐的首選地段，從火鍋、燒肉到中式料理與早午餐，每走幾步就有驚喜。以下是我實際造訪過的10間代表性餐廳清單，橫跨平價、創意、高級各路風格。

一頭牛日式燒肉｜炭香濃郁的和牛饗宴，約會聚餐首選

走在公益路上，很難不被 一頭牛日式燒肉 的木質外觀吸引。低調卻不失質感的門面，搭配昏黃燈光與暖色調的內裝，讓人一進門就感受到濃濃的日式職人氛圍。店內空間不大，但桌距規劃得宜，每桌皆設有獨立排煙設備，烤肉時完全不怕滿身油煙味。

餐點特色

一頭牛的靈魂，絕對是他們招牌的「三國和牛拼盤」。
嚴選的和牛部位，共八個部位、十樣餐點，讓人能從牛頭一路品嘗到牛尾。
油花分布均勻、切片厚薄恰好，經過炭火烤炙後香氣四溢，焦香與油脂在口中交融，入口即化的滑順感令人難忘。
值得一提的是，一頭牛的菜單設計十分彈性
想要一次體驗完整套餐也可以，偏好客製口味則能自由單點組合，不受套餐限制，想吃什麼就點什麼。
而且每桌都能選擇「自行燒烤」或「專人代烤」服務，烤肉管家的火侯掌握與節奏讓整體體驗更輕鬆愉快。
除了主角和牛，旬味野炊飯 與 主廚冰淇淋 也是隱藏版亮點，前者粒粒分明、香氣撲鼻；後者以香草與焙茶為基底，隨季節更換口味，完美收尾。整體服務親切熱情，特別是壽星還能享有 生日畫盤驚喜，讓慶祝時刻更添儀式感。

用餐體驗

整體節奏掌握得非常好。店員會在你剛想烤下一片肉時貼心遞上夾子、幫忙換烤網，讓人完全不用分心。整場用餐過程就像一場表演，從視覺、嗅覺到味覺都被滿足。
如果是第一次約會或慶祝特別節日，這裡的氛圍既不尷尬又不吵鬧，是營造氣氛的理想選擇。

綜合評分

地址：408臺中市南屯區公益路二段162號

電話：04-23206800

官網：http://www.marihuana.com.tw/yakiniku/index.html

小結語

一頭牛日式燒肉不僅是「吃肉的地方」，更像是一場五感盛宴。從進門那一刻到最後一道甜點，都能感受到他們對細節的用心。
若要在公益路找一間能讓人「邊吃邊微笑」的燒肉店，一頭牛 絕對值得列入你的必訪清單。

TANG Zhan 湯棧｜文青系火鍋代表，麻香湯底與視覺美感並重

在公益路這條美食戰線上，TANG Zhan 湯棧 是讓人一眼就會想走進去的那一種。
黑灰調的現代外觀、搭配微霧玻璃與招牌的「湯棧」燈字，呈現出一種低調的時尚感。
店內設計延續品牌主題，以「湯」為靈魂打造整體體驗，從裝潢到香氣，都有濃厚的溫潤氣息。

餐點特色

湯棧最有名的當然是它的「麻香鍋」。
湯底以雞骨與多種辛香料慢熬，香氣濃郁卻不嗆辣，入口後會在喉間留下柔和的花椒香。
「招牌麻油雞鍋」與「黃金牛奶鍋」也是人氣選項，特別是在冬天，溫潤的湯底配上滑嫩肉片，讓人每一口都覺得暖心。
他們的「滷肉飯」和「香蔥豆腐皮」更是許多老客人必點的靈魂配角，簡單卻有記憶點。

用餐體驗

整體氛圍比一般火鍋店更有質感。
桌距寬敞、燈光柔和，店員動作俐落又親切。即使客滿，也不會感覺吵雜或壓迫。
不論是一個人想靜靜吃鍋、或是朋友聚餐，湯棧都能給你剛剛好的距離與溫度。
值得一提的是，上菜速度快、湯底續湯毫不手軟，細節服務到位。

綜合評分

地址：408臺中市南屯區公益路二段248號

電話：04-22580617

官網：https://www.facebook.com/TangZhan.tw/

小結語

NINI 尼尼臺中店｜明亮寬敞的義式早午餐天堂

如果說前兩間是肉食愛好者的天堂，那 NINI 尼尼臺中店 絕對是想放鬆、聊聊天的好地方。餐廳外觀以白色系與大片玻璃窗為主，陽光灑進室內，讓人一踏入就有種度假般的輕盈感。假日早午餐時段特別熱鬧，建議提早訂位。

餐點特色

NINI 的菜單融合義式與臺灣人口味，選擇多樣且份量十足。主打的 松露燉飯 濃郁卻不膩口，米芯保留微Q口感；而 香蒜海鮮義大利麵 則以新鮮白蝦、花枝與淡菜搭配微辣蒜香，口感層次豐富。
此外，他們的薄餅披薩相當受歡迎，餅皮薄脆、餡料新鮮，是三五好友共享的好選擇。

用餐體驗

店內氣氛輕鬆不拘謹，無論是一個人帶電腦工作、或朋友聚餐，都能找到舒服角落。餐點上桌速度穩定，服務人員態度親切、補水與收盤都非常主動。整體節奏讓人覺得「時間變慢了」，很適合想遠離忙碌日常的人。

綜合評分

地址：40861臺中市南屯區公益路二段18號

電話：04-23288498

官網：https://nini.com.tw/

小結語

加分100%浜中特選昆布鍋物｜平價卻用心的湯頭系火鍋，家庭聚餐好選擇

在公益路這條高質感餐廳林立的戰場上，加分100%浜中特選昆布鍋物 走的是截然不同的路線。它沒有浮誇的裝潢、也沒有高價位的套餐，但靠著實在的湯頭與親切的服務，默默吸引許多回頭客。每到用餐時間，總能看到家庭或情侶三兩成群地圍著鍋邊聊天。

餐點特色

主打 北海道浜中昆布湯底，湯頭清澈卻不單薄，越煮越能喝出海藻與柴魚的自然香氣。
我這次點的是「牛奶昆布鍋」，入口時奶香與昆布香完美融合，搭配新鮮的牛五花肉片，滑順又不膩。
菜盤走健康取向，蔬菜比例高，連玉米、南瓜、豆皮都能吃出甜味；附餐的烏龍麵Q彈有嚼勁，吃完十分有飽足感。

用餐體驗

整體氛圍偏家庭取向，桌距寬敞、座位舒適，帶小孩來也不覺擁擠。店員態度親切，補湯、收盤都很勤快，給人一種「被照顧著」的安心感。
最難得的是，即使價位不高，食材新鮮度仍維持得很好，能感受到店家對品質的堅持。

綜合評分

地址：403臺中市西區公益路288號

電話：0910855180

官網：https://giafine100.com/

小結語

加分100%浜中特選昆布鍋物是一間「不浮誇、但會讓人想再訪」的火鍋店。它不追求豪華擺盤，而是用最簡單的湯頭與新鮮食材，傳遞出家常卻不平凡的溫度。
如果你想在公益路找一間可以放心帶家人一起吃的鍋物店，這裡絕對會讓人感到「加分」不少。

印月餐廳｜中式料理的藝術演繹，宴客與家庭聚會首選

說到臺中公益路的中式料理代表，印月餐廳 絕對是榜上有名。這間開業多年的餐廳以「中菜西吃」的概念聞名，把傳統中式料理以現代手法重新詮釋。從建築外觀到餐具擺設，每個細節都散發著低調的典雅氣息。
走進印月，挑高的空間、柔和的燈光與木質桌椅構成沉穩的氛圍。
不論是家庭聚餐、商務宴客，還是節日慶祝，都能找到恰到好處的格調。

餐點特色

印月最令人印象深刻的是他們將傳統中菜融入創意手法。
這次我品嚐的「松露雞湯」香氣濃郁、層次分明，一口下去既有中式的溫潤感，又帶出西式松露的奢華香氣。
「蒜香牛肋條」則是另一道招牌菜，外酥內嫩、油香十足，咬下去肉汁在口中散開，搭配特調醬汁非常過癮。
此外，他們的創意港點如「麻辣小籠包」與「金沙流沙包」也深受年輕客群喜愛，既保留經典又玩出新意。

用餐體驗

服務方面完全對得起餐廳的高級定位。從入座、點餐到上菜節奏，都拿捏得恰如其分。每道菜都會有服務人員細心介紹食材與吃法，讓人感受到「被款待」的尊榮感。
雖然價位偏中高，但在這樣的氛圍與品質下，物有所值。

綜合評分

地址：408臺中市南屯區公益路二段818號

電話：0422511155

官網：https://wein818.com/

小結語

印月餐廳是一間「不只吃飯，更像品味生活」的地方。
它成功地讓中式料理不再只是圓桌菜，而是能展現質感、講究細節的美食體驗。
若你在找一間能同時滿足味蕾與體面的餐廳，印月 絕對是公益路上的不敗經典。

KoDō 和牛燒肉｜極致職人精神，專為儀式感與頂級味覺而生

若要形容 KoDō 和牛燒肉 的用餐體驗，一句話足以總結——「像在欣賞一場關於肉的表演」。
隱身在公益路一隅，KoDō 的外觀低調典雅，店內以深色木質調與間接照明營造出沉穩氛圍。
從踏入店門那一刻開始，服務人員的態度、動線、聲音控制，全都精準到位，讓人彷彿走進日式劇場。

餐點特色

這裡主打 日本A5和牛冷藏肉，以「精切厚燒」的方式呈現。
我點的「壽喜燒風和牛套餐」是本日最驚艷的一道——服務人員現場以鐵鍋輕煎，再淋上特製壽喜燒醬汁，香氣瞬間瀰漫整桌。
肉片油花細緻、入口即化，搭配生蛋液後更添柔滑口感。
另一道「冷藏肋眼心」則保留了和牛的彈性與甜度，每一口都能感受到油脂與炭火交織出的層次。
即使是配角如「季節小菜」與「日式和風飯」也毫不馬虎，整體呈現出高級卻不造作的平衡。

用餐體驗

KoDō 的最大特色是「儀式感」。
每位店員的動作都有節奏，從擺盤、火候、換網到講解，都像排練過無數次的演出。
在這裡用餐，會自然地放慢速度，專注於每一口肉帶來的細膩變化。
特別推薦搭配店內的紅酒或日本威士忌，風味更加圓潤。

綜合評分

地址：403臺中市西區公益路260號

電話：0423220312

官網：https://www.facebook.com/kodo2018/

小結語

KoDō 和牛燒肉不是日常餐廳，而是一場體驗。
從環境、服務到食材，每個細節都讓人感受到對「完美」的執著。
若你想在公益路找一間能讓人留下深刻印象、適合紀念日慶祝的餐廳，KoDō 絕對是值得收藏的一次「味覺儀式」。

永心鳳茶｜在茶香裡用餐的優雅時光，臺味早午餐的新詮釋

走進 永心鳳茶公益店，彷彿進入一間有氣質的茶館。
柔和的燈光灑在復古綠牆上，搭配大理石桌面與金色餐具，整體氛圍既典雅又帶有一絲文青氣息。
這裡不只是喝茶的地方，更像是把「臺灣味」以早午餐的形式重新演繹。

餐點特色

永心鳳茶的餐點結合中式靈魂與西式擺盤，無論是「炸雞腿飯」還是「紅玉紅茶拿鐵」，都能讓人感受到熟悉卻不平凡的味道。
炸雞腿外酥內嫩，搭配自製酸菜與溏心蛋，鹹香中帶著層次感。
「鳳茶甜點拼盤」則以茶為靈魂——伯爵茶蛋糕、烏龍茶奶酪、紅茶雪酥，每一口都有細緻的香氣變化。
最特別的是他們的茶飲，從臺灣高山紅茶到金萱冷泡茶，每一壺都現泡現倒，香氣清雅。
對我而言，這不只是一頓飯，更是一段放鬆的午後儀式。

用餐體驗

店內服務人員態度溫和，對茶品介紹詳盡。上餐節奏剛好，不急不徐。
整體氛圍很「耐坐」，許多客人吃完正餐後仍會續點一壺茶聊天。
音樂輕柔、光線柔和，是那種可以靜靜待上兩小時的地方。

綜合評分

地址：40360臺中市西區公益路68號三樓(勤美誠品)

電話：0423221118

官網：https://linktr.ee/yonshin

小結語

永心鳳茶讓人重新定義「臺味」。
它不走傳統路線，而是把熟悉的元素以更細緻、更現代的方式呈現。
無論是姊妹下午茶、親子餐聚，或是想一個人沉澱片刻，永心鳳茶 都是一處能讓人慢下來、品味生活的好地方。

三希樓｜老饕級江浙功夫菜，穩重又帶人情味的中式饗宴

位於公益路上的 三希樓 是許多臺中老饕的口袋名單。
它沒有浮誇的裝潢，卻有一種低調的自信。從大門進入，就能聞到淡淡的醬香與蒸氣味，那是正宗江浙菜的靈魂。
整體裝潢以深木色為主，搭配圓桌與包廂設計，非常適合家庭聚餐或請客宴會。

餐點特色

三希樓的菜色以 江浙與港式料理 為主，兼顧傳統與現代風味。
我這次點了「東坡肉」與「蝦仁炒飯」，兩道都展現了主廚深厚的火候功力。
東坡肉油亮卻不膩，入口即化、鹹甜交織；蝦仁炒飯粒粒分明、香氣十足，每一口都吃得到鑊氣。
此外，「小籠包」皮薄多汁，是幾乎每桌必點的招牌；港點類如「金牌流沙包」與「干貝燒賣」也都表現穩定。

用餐體驗

三希樓的服務給人一種老派但貼心的感覺。
店員上菜節奏掌握得很好，會主動幫忙分菜、收盤，態度沉穩而不打擾。
最讓我印象深刻的是，這裡的客群非常多元——有帶長輩的家庭、公司聚餐，也有情侶共度節日，卻都能在同一空間裡感到自在。

綜合評分

地址：408臺中市南屯區公益路二段95號

電話：0423202322

官網：https://www.sanxilou.com.tw/

小結語

三希樓是一間「吃得出功夫」的餐廳。
它不追求創新，而是用穩定的味道與真材實料，抓住每一位饕客的胃。
如果你想在公益路上找一間能兼顧長輩口味、氣氛又不拘謹的中餐廳，三希樓 絕對是最穩妥的選擇。

一笈壽司｜低調奢華的無菜單日料，職人手藝詮釋旬味極致

在熱鬧的公益路上，一笈壽司 低調得幾乎不顯眼。
外觀簡約，沒有華麗招牌，只有小小的木質門面與柔黃燈光。
一推開門，迎面而來的是日式杉木香氣與寧靜的氛圍，吧檯座位整齊排列，主廚站在中間，彷彿舞臺上的演出者。

餐點特色

一笈壽司採 Omakase（無菜單料理） 形式，每一餐都由主廚根據當日食材設計。
我這次選擇中價位套餐（約 $1200），共十多道料理，從前菜、小鉢、刺身、握壽司到甜點一氣呵成。
「比目魚鰭邊握」是整場最驚豔的瞬間——主廚以火槍輕炙，油脂瞬間釋放，入口後化成柔滑香氣。
「甜蝦海膽軍艦」則完美展現鮮度與層次感，海膽甘甜、甜蝦緊實。
搭配主廚親自調配的醬汁，每一口都像在品嚐季節的節奏。

用餐體驗

整場用餐約90分鐘，節奏緩慢但沉穩。
主廚會邊料理邊與客人互動，介紹魚種產地與食材處理方式。
雖然整體空間不大，但氣氛極佳——柔和的音樂、清酒的香氣、刀刃切魚時的聲音，讓人完全沉浸其中。
特別喜歡他們最後的甜點「焙茶奶酪」，收尾清爽優雅，為整場體驗畫下完美句點。

綜合評分

地址：408臺中市南屯區公益路二段25號

電話：0423206368

官網：https://www.facebook.com/YIJI.sushi/

小結語

一笈壽司是一間真正讓人「放慢呼吸」的餐廳。
這裡沒有多餘擺盤，也不靠噱頭，而是以主廚對食材的尊重與技術堆疊出一場味覺饗宴。
若你想在公益路體驗日本料理最純粹的精神，一笈壽司 絕對值得你預約、靜靜期待。

茶六燒肉堂｜人氣爆棚的和牛燒肉聖地，肉香與幸福感同時滿分

若要票選公益路上「最難訂位」的餐廳，茶六燒肉堂 絕對名列前茅。
不管平日或假日，用餐時段幾乎一位難求。外觀以木質格柵搭配大面玻璃設計，呈現出年輕又有質感的風格。店內空間明亮、桌距適中，播放著輕快的音樂，整體氛圍熱鬧中帶點高級感，是許多年輕人聚餐、慶生的首選地。

餐點特色

茶六主打 和牛燒肉套餐，價格約落在 $700–$1000 間，份量與品質兼具。
我這次點的是「厚切牛舌套餐」，肉片厚實彈牙，略帶脆感，搭配鹽蔥提味剛剛好。
另一道「和牛拼盤」也相當受歡迎，油花分布均勻、香氣濃郁，輕烤幾秒即可入口即化。
套餐附餐部分也相當用心：沙拉新鮮、味噌湯濃郁，最後還有一份「茶香冰淇淋」作結尾，香氣清爽，完美收尾。

用餐體驗

茶六的服務效率相當高。店員親切、換網勤快、補水速度快，整場用餐流程流暢無壓力。
雖然客人很多，但環境維持得乾淨整潔，動線規劃良好。
最令人印象深刻的是他們的 整體節奏拿捏得剛剛好 ——餐點上桌快、氣氛熱絡，卻不會讓人覺得匆忙。
不論是朋友聚會、家庭聚餐，甚至是情侶約會，都能找到各自的樂趣。

綜合評分

地址：403臺中市西區公益路268號

電話：0423281167

官網：https://inline.app/booking/-L93VSXuz8o86ahWDRg0:inline-live-karuizawa/-LUYUEIOYwa7GCUpAFWA

小結語

茶六燒肉堂用「穩定品質＋輕奢氛圍」抓住了臺中年輕族群的心。
不論是第一次約會還是老朋友重聚，都能在這裡找到屬於燒肉的快樂節奏。
若你在公益路只想挑一家「保證不踩雷」的燒肉店，茶六燒肉堂 絕對是首選。

吃完10家公益路餐廳後的心得與結語

吃完這十家餐廳後，臺中公益路不只是一條美食街，而是一段生活風景線。

有的餐廳講究細膩與儀式感，像 一頭牛日式燒肉 與 一笈壽司，讓人感受到食材最純粹的美好

有的則以親切與溫度打動人心，像 加分昆布鍋物、永心鳳茶，讓人明白吃飯不只是為了飽足，而是一種被照顧的幸福。

而像茶六燒肉堂、TANG Zhan 湯棧 這類人氣名店，則用穩定的品質與熱絡的氛圍，成為許多臺中人心中「想吃肉就去那裡」的代名詞。

這十家店，構成了公益路最動人的縮影

有華麗的，也有溫柔的；有傳統的，也有創新的。

 每一家都在自己的風格裡發光，讓人吃到的不只是料理，而是一種生活的溫度與節奏。

對我而言，這不僅是一場美食旅程，更是一趟關於「臺中味道」的回憶之旅。

FAQ：關於臺中公益路美食常見問題

Q1：公益路哪一區的餐廳最集中？
 最熱鬧的區段大約在「公益路與黎明路口」一帶，這裡聚集了許多知名餐廳，從高級燒肉到早午餐通通有。
像 一頭牛日式燒肉、TANG Zhan 湯棧、茶六燒肉堂 都在這附近，交通方便、停車也相對容易。

Q2：需要提前訂位嗎？
 公益路的熱門餐廳幾乎都建議 提早3～5天訂位，尤其是假日或節慶期間。
特別是 一頭牛日式燒肉、KoDō 和牛燒肉、一笈壽司 這幾家，若臨時前往幾乎很難有位。

最後的話

若要用一句話形容這趟美食之旅，我會說：
「在公益路，吃飯不是選擇，而是一種享受。」
這條路上的每一次用餐，都像一段城市裡的小旅行。
下次當你不確定想吃什麼時，不妨沿著公益路走一圈，或許下一家，正好就是你新的最愛。

NINI 尼尼臺中店有雷嗎？

如果你也和我一樣喜歡用味蕾探索一座城市，那就把這篇公益路美食攻略收藏起來吧。NINI 尼尼臺中店份量足夠嗎？

無論是約會、慶生、家庭聚餐，或只是想犒賞一下辛苦的自己——這條路上永遠會有一間剛剛好的餐廳在等你。一頭牛日式燒肉年末聚餐推薦嗎？

下一餐，不妨從這10家開始。永心鳳茶服務態度如何？

打開手機、約上朋友，讓公益路成為你生活裡最容易抵達的小確幸。加分100%浜中特選昆布鍋物好吃嗎？

如果你有私心愛店，也歡迎留言分享，TANG Zhan 湯棧人潮很多嗎？

你的推薦，可能讓我下一趟美食旅程變得更精彩。加分100%浜中特選昆布鍋物有雷嗎？

A new study suggests that the weathering of sulfate rocks, not increasing ocean phosphorus levels, was crucial to the oxygenation of Earth’s atmosphere, influencing the late evolution of animal life, and also indicates that potential for complex intelligent life on other planets might require longer incubation times. Scientists identified sulfate as the missing link in Earth’s oxygenation, reshaping our understanding of life’s evolution and the potential for intelligent life on other planets. A recent research study may have discovered a missing link that helps explain Earth’s uniquely oxygen-saturated atmosphere and the corresponding evolution of animal life on our planet. The study, led by a Fellow of the Forrest Research Foundation at The University of Western Australia and recently published in the esteemed journal Nature, may hold the key to understanding why, for almost 90% of Earth’s history, oxygen levels remained too low for animals to breathe. The first major evolutionary event of animal life occurred during an event dubbed the Shuram Excursion – between 570 and 550 million years ago – which is believed to represent a massive release of carbon dioxide and oxygen into the atmosphere and oceans as a result of increasing ocean phosphorus levels. To test the theory, researchers used a newly developed tool to track the abundance of phosphorus in the oceans hundreds of millions of years ago, recorded in six locations in Australia, China, Mexico, and the US. Sulfate: The True Driver of Earth’s Oxygenation? The data and Earth chemistry model revealed increasing ocean phosphorus levels could not have explained the rise of oxygen. The effect was only replicated by the model when large quantities of sulfate rock were weathered, releasing sulfate into the oceans to produce vast amounts of oxygen. Lead author and Forrest Fellow Dr. Matthew Dodd, from the UWA School of Earth Sciences, said the results suggested sulfate, rather than phosphorus, was the main control in the oxygenation of the planet during the first major evolution of complex life. “Our findings may explain the prolonged low levels of oxygen throughout Earth’s history and consequently the late evolution of animal life on Earth,” Dr. Dodd said. “Importantly, we observe that ocean phosphorus was predominantly low when oxygen levels were low throughout the Shuram Excursion. This phenomenon would have locked the early oceans and atmosphere into an oxygen-devoid state.” Data from the study also holds implications about the possibility of intelligent life on other planets. “These results suggest other potentially habitable planets may support complex intelligent life, only if provided with long enough incubation times,” Dr. Dodd said. “This could imply planets around stars larger than the Sun may not develop complex intelligent life due to the relatively short lifetime of large stars.” Reference: “Uncovering the Ediacaran phosphorus cycle” by Matthew S. Dodd, Wei Shi, Chao Li, Zihu Zhang, Meng Cheng, Haodong Gu, Dalton S. Hardisty, Sean J. Loyd, Malcolm W. Wallace, Ashleigh vS. Hood, Kelsey Lamothe, Benjamin J. W. Mills, Simon W. Poulton and Timothy W. Lyons, 31 May 2023, Nature. DOI: 10.1038/s41586-023-06077-6

Bamboo longhorn beetle (Chlorophorus annularis) captured in Braintree, United Kingdom. Credit: Stephen Rolls The Asian bamboo longhorn beetle is more widespread in Europe than previously known. A worryingly high number of Asian bamboo longhorn beetles (Chlorophorus annularis) turn out to have been emerging across Europe for about a century already, finds an international research team, headed by researchers from the Center of Natural History, University of Hamburg, Germany. Curiously, the recent records of the invasive, non-native to the Old Continent species are mostly sourced from citizen scientists and online platforms, which proves the power of involving the public in species monitoring. The study is published in the open-access, peer-reviewed scientific journal BioRisk. In our globalized world, which has already become victim to climate change and biodiversity loss, non-native species present a further threat to our ecosystems. Thus, the rising accounts of newly recorded alien species are of serious concern to both scientists and (inter)national institutions. However, surveying non-native species remains limited to a small fraction of species: those known to be particularly invasive and harmful. Emerging Asian bamboo longhorn beetle from bamboo in the Czech Republic. Credit: Petr Jansa Global Trade Fuels the Beetle’s Silent Spread One of the multitude of non-native species that are currently lacking efficient and coordinated surveying efforts is the Asian bamboo longhorn beetle (Chlorophorus annularis). Naturally occurring in temperate and tropical Southeast Asia, the insect feeds on a variety of plants, but prefers bamboo. Thus, due to the international trade of bamboo and the insects ‘traveling’ with the wood, the species has continuously been expanding its distribution around the world. Its first appearance in Europe was recorded back in 1924, when it was identified in England. Bamboo longhorn beetle (Chlorophorus annulari) captured in Lincoln, United Kingdom. Credit: Sheena Cotter Back to our days, during a fieldwork practice for students at the University of Hamburg, held within the city because of the COVID-19 traveling restrictions, the team stumbled across a longhorn beetle, later identified by scientists as the Asian bamboo borer. Furthermore, it became clear that there were even more recent records published across different citizen science platforms, such as iNaturalist, iRecord and Waarneming.nl. Having taken the contacts of the citizen scientists from there, the researchers approached them to ask for additional collection details and images, which were readily provided. As a result, the researchers formally confirmed the presence of the Asian bamboo borer in Belgium and the Netherlands. In total, they reported thirteen new introductions of the species in Europe, which translates to a 42% increase in the records of the species for the continent. Climate Change and Garden Trends May Aid Establishment “In light of the warming climate and a growing abundance of ornamental bamboo plants in Europe, the beetle might get permanently established. Not only could it become a garden pest, but it could also incur significant costs to the bamboo-processing industry,” comments Dr. Matthias Seidel, lead author of the study. Having realized the potential of citizen science for bridging the gaps in invasive species monitoring, the researchers now propose for specialized platforms to be established with the aim to familiarise non-professional scientists with non-native species of interest and provide them with more sophisticated reporting tools. The aim is to speed up the identification of important alien species by collating records of specific species of interest, which are flagged and regularly exported from other citizen science databases and platforms. Reference: “Citizen scientists significantly improve our knowledge on the non-native longhorn beetle Chlorophorus annularis (Fabricius, 1787) (Coleoptera, Cerambycidae) in Europe” by Matthias Seidel, Maren Lüttke, Christian Cocquempot, Katy Potts, Wil J. Heeney and Martin Husemann, 9 March 2021, BioRisk. DOI: 10.3897/biorisk.16.61099

Mammalian cells that have been successfully genetically engineered using the STAMPScreen method. Credit: Wyss Institute at Harvard University STAMPScreen Pipeline Helps Streamline Genetic Studies in Mammalian Cells Today’s genetic engineers have a plethora of resources at their disposal: an ever-increasing number of massive datasets available online, highly precise gene editing tools like CRISPR, and cheap gene sequencing methods. But the proliferation of new technologies has not come with a clear roadmap to help researchers figure out which genes to target, which tools to use, and how to interpret their results. So, a team of scientists and engineers at Harvard’s Wyss Institute for Biologically Inspired Engineering, Harvard Medical School (HMS), and the MIT Media Lab decided to make one. The Wyss team has created an integrated pipeline for performing genetic screening studies, encompassing every step of the process from identifying target genes of interest to cloning and screening them quickly and efficiently. The protocol, called Sequencing-based Target Ascertainment and Modular Perturbation Screening (STAMPScreen), is described in Cell Reports Methods, and the associated open-source algorithms are available on GitHub. The STAMPScreen workflow is an integrated pipeline that allows researchers to quickly and easily analyze an experimental database for potential genes of interest (1), choose which screening tool to use (2), create a screening library (3), and use next-generation sequencing to screen genes in vivo (4). The individual steps can also be used in other workflows. Credit: Wyss Institute at Harvard University “STAMPScreen is a streamlined workflow that makes it easy for researchers to identify genes of interest and perform genetic screens without having to guess which tool to use or what experiments to perform to get the results they want,” said corresponding author Pranam Chatterjee, Ph.D., a former graduate student at the MIT Media Lab who is now the Carlos M. Varsavsky Research Fellow at HMS and the Wyss Institute. “It is fully compatible with many existing databases and systems, and we hope that many scientists are able to take advantage of STAMPScreen to save themselves time and improve the quality of their results.” Frustration is the mother of invention Chatterjee and Christian Kramme, a co-first author of the paper, were frustrated. The two scientists were trying to explore the genetic underpinnings of different aspects of biology — like fertility, aging, and immunity — by combining the strengths of digital methods (think algorithms) and genetic engineering (think gene sequencing). But they kept running into problems with the various tools and protocols they were using, which are commonplace in science labs. The algorithms that purported to sift through an organism’s genes to identify those with a significant impact on a given biological process could tell when a gene’s expression pattern changed, but didn’t provide any insight into the cause of that change. When they wanted to test a list of candidate genes in living cells, it wasn’t immediately clear what type of experiment they should run. And many of the tools available to insert genes into cells and screen them were expensive, time-consuming, and inflexible. Co-first author of the paper, Christian Kramme, at his bench at the Wyss Institute. Credit: Wyss Institute at Harvard University “I was using methods known as Golden Gate and Gateway to clone genes into vectors for screening experiments, and it took me months and thousands of dollars to clone 50 genes. And using Gateway, I couldn’t physically barcode the genes to identify which one got into which vector, which was a crucial requirement for my downstream sequencing-based experimental design. We figured there had to be a better way to do this kind of research, and when we couldn’t find one, we took on the challenge of creating it ourselves,” said Kramme, who is a graduate student at the Wyss Institute and HMS, Kramme teamed up with co-first author and fellow Church lab member Alexandru Plesa, who was experiencing identical frustrations making gene vectors for his project. Kramme, Plesa, and Chatterjee then set to work outlining what would be required to make an end-to-end platform for genetic screening that would work for all of their projects, which ranged from protein engineering to fertility and aging. From bits to the bench To improve the earliest stage of genetic research — identifying genes of interest to study — the team created two new algorithms to help meet the need for computational tools that can analyze and extract information from the increasingly large datasets that are being generated via next-generation sequencing (NGS). The first algorithm takes the standard data about a gene’s expression level and combines it with information about the state of the cell, as well as information about which proteins are known to interact with the gene. The algorithm gives a high score to genes that are highly connected to other genes and whose activity is correlated with large, cell-level changes. The second algorithm provides more high-level insight by generating networks to represent the dynamic changes in gene expression during cell-type differentiation and then applying centrality measures, such as Google’s PageRank algorithm, to rank the key regulators of the process. MegaGate, a novel method for cloning target genes of interest into vectors, is much more efficient at producing successful gene-bearing vectors (left) than other existing methods like Gateway (right). Credit: Wyss Institute at Harvard University “The computational part of genetic studies is like a Jenga game: if each block in the tower represents a gene, we’re looking for the genes that make up the base of the Jenga tower, the ones that hold the whole thing up. Most algorithms can only tell you which genes are in the same row as each other, but ours allow you to home in on how far up or down the tower they are, so you can quickly identify the ones that have the biggest influence on the cell state in question,” said Chatterjee. Once the target genes have been identified, the STAMPScreen protocol moves from the laptop to the lab, where experiments are performed to disrupt those genes in cells and see what effect that perturbation has on the cell. The team of researchers systematically evaluated multiple gene perturbation tools including complementary DNA (cDNA) and several versions of CRISPR in human induced pluripotent stem cells (hiPSCs), the first known head-to-head comparisons performed entirely in this highly versatile yet challenging cell type. They then created a new tool that allows CRISPR and cDNA to be used within the same cell to unlock synergies between the two methods. For example, CRISPR can be used to turn off expression of all isoforms of a gene, and cDNA can be used to sequentially express each isoform individually, allowing more nuanced genetic studies and greatly reducing background expression of off-target genes. Scanning library barcodes The next step in many genetic experiments is generating a screening library for introducing genes into cells and observing their effects. Typically, gene fragments are inserted into bacterial plasmids (circular pieces of DNA) using methods that work well for small pieces of DNA, but are cumbersome to use when inserting larger genes. Many of the existing methods also rely on a technique called Gateway, which uses a process called lambda phage recombination and the production of a toxin to kill off any bacteria that did not receive a plasmid with the gene of interest. The toxin in these plasmids is often cumbersome to work with in the lab, and can be inadvertently inactivated when a “barcode” sequence is added to a vector to help researchers identify which gene-bearing plasmid the vector received. Kramme and Plesa were working with Gateway when they realized that these problems could be solved if they eliminated the toxin and replaced it with short sequences on the plasmid that would be recognized and cut by a type of enzyme called meganucleases. Meganuclease recognition sequences do not appear in the genes of any known organism, thus ensuring that the enzyme will not accidentally cut the inserted gene itself during cloning. These recognition sequences are naturally lost when a plasmid receives a gene of interest, making those plasmids immune to meganuclease. Any plasmids that do not successfully receive the gene of interest, however, retain these recognition sequences and are cut to pieces when a meganuclease is added, leaving only a pure pool of plasmids containing the inserted gene. The new method, which the researchers dubbed MegaGate, had a cloning success rate of 99.8% and also allowed them to barcode their vectors with ease. “MegaGate not only solves many of the problems that we kept running into with older cloning methods, it is also compatible with many existing gene libraries like the TFome and hORFeome. You can essentially take Gateway and meganucleases off the shelf, put them together with a library of genes and a library of barcoded destination vectors, and two hours later you have your barcoded genes of interest. We’ve cloned nearly 1,500 genes with it, and have yet to have a failure,” said Plesa, who is a graduate student at the Wyss Institute and HMS. Finally, the researchers demonstrated that their barcoded vectors could be successfully inserted into living hiPSCs, and pools of cells could be analyzed using NGS to determine which delivered genes were being expressed by the pool. They also successfully used a variety of methods, including RNA-Seq, TAR-Seq, and Barcode-Seq, to read both the genetic barcodes and the entire transcriptomes of hiPSCs, enabling researchers to use whichever tool they are most familiar with. The team anticipates that STAMPScreen could prove useful for a wide variety of studies, including pathway and gene regulatory network studies, differentiation factor screening, drug and complex pathway characterizations, and mutation modeling. STAMPScreen is also modular, allowing scientists to integrate different parts of it into their own workflows. “There’s a treasure trove of information housed in publicly available genetic datasets, but that information will only be understood if we use the right tools and methods to analyze it. STAMPScreen will help researchers get to eureka moments faster and speed up the pace of innovation in genetic engineering,” said senior author George Church, Ph.D., a Wyss Core Faculty member who is also a Professor of Genetics at HMS and Professor of Health Sciences and Technology at Harvard and MIT. “At the Wyss Institute we aim for impactful ‘moonshot’ solutions to pressing problems, but we know that to get to the moon, we have to first build a rocket. This project is a great example of how our community innovates on-the-fly to enable scientific breakthroughs that will change the world for the better,” said Wyss Founding Director Don Ingber, M.D., Ph.D., who is also the Judah Folkman Professor of Vascular Biology at HMS and the Vascular Biology Program at Boston Children’s Hospital, as well as Professor of Bioengineering at Harvard John A. Paulson School of Engineering and Applied Sciences. Reference: “An Integrated Pipeline for Mammalian Genetic Screening” by Christian Kramme, Alexandru M. Plesa, Helen H. Wang, Bennett Wolf, Merrick Pierson Smela, Xiaoge Guo, Richie E. Kohman, Pranam Chatterjee and George M. Church, 27 September 2021, Cell Reports Methods. DOI: 10.1016/j.crmeth.2021.100082 Additional authors of the paper include Helen Wang, Bennett Wolf, Merrick Smela, Xiaoge Guo, Ph.D., and Richie Kohman, Ph.D. from the Wyss Institute and HMS.

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